Device for screening medicine for resisting colorectal cancer pulmonary metastasis

Abstract

The utility model belongs to the field of medical instruments, particularly relates to a device for screening medicines for resisting colorectal cancer pulmonary metastasis, and provides the following scheme aiming at the problem that a plurality of links of apoptosis resistance, liquid-phase shearing force resistance and the like of cancer cells in blood flow cannot be simulated in the prior art: the device comprises a screening mechanism, the screening mechanism comprises a liquid-phase flowing assembly and a plurality of diversion grooves, the diversion grooves and the liquid-phase flowing assembly form a closed circulating flowing system, the colorectal cell culture medium serving as a flowing liquid phase circularly flows in the system, and colorectal cancer cells circularly flow in the liquid phase and interact with lung organs. The condition is close to the in-vivo condition; a plurality of processes of cancer cell metastasis are simulated in vitro, the drug can be sensitively reflected to a fluorescence intensity result as long as the drug can generate an intervention effect on one of the links, and screening of a plurality of anti-metastasis new drugs is facilitated; the method belongs to an in-vitro screening model and is simple to operate.

Description

Technical Field

[0001] The utility model relates to a drug screening device, in particular to a device for screening drugs for resisting colorectal cancer lung metastasis, and belongs to the technical field of medical devices. Background Art

[0002] Colorectal cancer is the third most common cancer in the world, posing a serious threat to human health and life. In 2020 alone, there were more than 1.93 million new cases and more than 930,000 deaths worldwide. The main cause of treatment failure and death in colorectal cancer patients is not the primary lesion, but distant metastasis. Lung metastasis is the most common extra-abdominal metastasis of colorectal cancer, accounting for 32.9% of all colorectal cancer metastasis cases. Once lung metastasis occurs, the patient's prognosis is extremely poor, with a median survival time of only 8 months, a 1-year survival rate of only 30%, and a 5-year survival rate of only 5%. The development of drugs to prevent and treat lung metastasis is the key to prolonging the survival of colorectal cancer patients. However, progress in the development of such drugs has been very slow. The key reason is that there is no suitable in vitro screening model, and large-scale in vitro drug screening cannot be carried out.

[0003] Pulmonary metastasis of colorectal cancer is a complex process, including cancer cells falling off from the primary lesion, entering the blood circulation, escaping apoptosis in the blood, migrating to the lung tissue under the action of lung chemokines, degrading the extracellular matrix of the lung, invading the lung tissue, and proliferating in the lung tissue. Although the nude mouse transplant tumor model can reproduce this process more completely, the construction of the nude mouse orthotopic transplant tumor model of colon cancer is difficult, time-consuming, labor-intensive, and expensive. In addition, the probability of liver metastasis is not high, and the metastasis between individuals and batches varies greatly, making it difficult to use for large-scale drug screening. As an in vitro model, the Transwell invasion model is often used to study the metastasis and invasion of cancer cells. However, this model is a static model that can only reflect the link of cancer cells penetrating the basement membrane. It cannot simulate the pathological process of cancer cells resisting apoptosis and resisting liquid shear force in the blood flow, and is not suitable for efficient screening of drugs against colon cancer lung metastasis. Therefore, there is an urgent need for a new device that can effectively simulate the pathological process of colorectal cancer lung metastasis and can be easily operated in vitro with stable effects for new drug screening. Summary of the invention

[0004] The purpose of the utility model is to provide a device for screening drugs against colorectal cancer lung metastasis in order to solve at least one of the above technical problems.

[0005] The utility model achieves the above-mentioned purpose through the following technical scheme: a device for screening drugs against colorectal cancer lung metastasis, comprising a screening mechanism, a screening component for screening drugs against colorectal cancer lung metastasis is placed on a frame mechanism, and the screening mechanism comprises a liquid phase flow component and a plurality of diversion troughs;

[0006] The diversion tank includes a bottom tank and a screening plate, the screening plate is placed in the bottom tank, a plurality of culture recesses are arranged on the screening plate, a guide plate is connected to the upper plate surface at one end of the screening plate, and both ends of the bottom tank are connected to a docking port;

[0007] The liquid phase flow component includes a circulation pump and a delivery pipeline. The inlet and outlet liquid ends of the circulation pump are connected to the delivery pipeline, and the delivery pipeline is also connected to the two ends of the diversion trough. A liquid storage tank is arranged on the delivery pipeline, and the liquid storage tank is provided with an impurity removal component consisting of an adsorption unit, a stirring unit and a power unit.

[0008] As a further solution of the utility model: the frame mechanism includes an upper support plate, a lower support plate and a column, the upper support plate and the lower support plate are distributed in parallel up and down, the column is vertically connected to the four corners of the upper support plate and the lower support plate, a diversion trough is arranged on the upper support plate, and a liquid flow component is fixedly connected to the lower support plate.

[0009] As a further solution of the utility model: a limiting groove is provided at the center of the upper plate surface of the upper support plate, a plurality of magnets are embedded in the bottom surface of the limiting groove, and an iron sheet is embedded in the bottom surface of the bottom groove.

[0010] As a further solution of the utility model: a liquid inlet joint group and a liquid outlet joint group are respectively provided on the short sides of both sides of the upper support plate, the bottom ends of the liquid inlet joint group and the liquid outlet joint group are connected to the conveying pipeline, and the upper ends of the liquid inlet joint group and the liquid outlet joint group are connected to the docking port through a connecting hose.

[0011] As a further solution of the utility model: a reserved hole is opened on the bottom surface of the bottom groove, and the reserved holes are arranged in a one-to-one correspondence with the culture recessed holes, and organoids of normal human lung tissue embedded in Matrigel are cultured in the culture recessed holes.

[0012] As a further solution of the utility model: a plurality of guide plates are arranged at one end of the screening plate close to the liquid inlet joint group, and the guide plates are arranged in a trumpet shape.

[0013] As a further solution of the utility model: the adsorption unit of the impurity removal component includes a impurity collecting pipe composed of an impurity collecting shell and an adsorption filter element. The impurity collecting pipe is detachably inserted vertically in the liquid storage tank. The impurity collecting pipe is tightly attached to the inner wall of the liquid storage tank, and the adsorption filter element is filled in the impurity collecting shell.

[0014] As a further solution of the utility model: the stirring unit of the impurity removal component includes a turntable and a flip plate, the turntable is rotatably connected to the bottom end of the liquid storage tank, the flip plate is connected to the turntable in a cross shape, and the upper end of the flip plate is inclined in the rotation direction.

[0015] As a further solution of the utility model: the power unit of the impurity removal component includes a driving motor and a driving wheel. The driving motor is fixedly connected to the U-shaped frame in an inverted state. The U-shaped frame is fixedly welded to the lower support plate. The driving wheel is coaxially arranged with the turntable through a connecting shaft, and the rotating shaft of the driving motor is connected to the driving wheel through a transmission belt.

[0016] As a further solution of the utility model: the side wall of the liquid storage tank is connected to a delivery pipeline for inputting the flowing liquid phase, the upper end of the liquid storage tank is connected to a delivery pipeline for outputting the flowing liquid phase, and the delivery pipeline for outputting the flowing liquid phase is inserted into the bottom of the liquid storage tank.

[0017] The beneficial effects of the utility model are:

[0018] 1. The flow trough and the liquid phase flow component form a closed circulation flow system. The colorectal cell culture medium as the mobile liquid phase circulates in the system. The colorectal cancer cells circulate in the liquid phase and interact with the lung organoids, which is closer to the in vivo situation.

[0019] 2. Multiple shunt slots are provided, and culture recesses are provided on the screening plate provided in the shunt slots. Organoids of normal human lung tissue embedded in Matrigel can be cultured in the culture recesses of each screening plate, ensuring that the screening plate can be placed horizontally on the bottom surface of the bottom slot. During the growth process, lung organoids will release factors specific to lung tissue, which will have a chemotactic effect on colorectal cancer cells in the mobile liquid phase. Colorectal cancer cells with metastatic ability will move toward the recesses, adhere to the surface of Matrigel, penetrate Matrigel, enter the gel droplets, interact with lung organoids, proliferate and form clones. The fluorescence intensity in the gel droplets can be detected under a live cell imaging instrument to determine the metastatic ability of metastatic colorectal cancer cells. Multiple processes of cancer cell metastasis can be simulated in vitro. As long as the drug can have an intervention effect on one of the links, it can be sensitively reflected in the fluorescence intensity results, which is conducive to screening a variety of new anti-metastatic drugs;

[0020] 3. An impurity removal component is provided to drive the liquid phase in the tank to rotate, so that the impurities in the liquid phase are moved to the tank wall under the action of centrifugal force. The impurities contained in the liquid phase can be adsorbed by the adsorption filter element to the maximum extent, so that the flowing liquid phase entering the diversion tank again can be free of impurities;

[0021] 4. A guide plate is connected to the screening plate, so that the flowing liquid phase entering the bottom tank can flow evenly on the screening plate under the action of the guide plate;

[0022] 5. This device is an in vitro screening model, which is low-cost and easy to operate compared with animal models. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a schematic diagram of the overall structure of the utility model;

[0024] Figure 2 This is a schematic diagram of the split structure of the diverter trough of the utility model;

[0025] Figure 3 This is a schematic diagram of the upper support plate structure of the utility model;

[0026] Figure 4 This is a schematic diagram of the bottom structure of the diverter trough of the utility model;

[0027] Figure 5 This is a schematic diagram of the connection structure between the liquid storage tank and the drive motor of the utility model;

[0028] Figure 6 This is a schematic diagram of the cross-sectional structure of the liquid storage tank of the utility model;

[0029] Figure 7 This is a schematic diagram of the structure of the adsorption unit of the utility model;

[0030] Figure 8 It is a schematic diagram of the structure of the handover unit of the utility model.

[0031] In the figure: 1. upper support plate, 2. lower support plate, 3. column, 4. diversion trough, 41. bottom trough, 42. screening plate, 43. upper cover, 44. reserved hole, 45. docking port, 46. culture recessed hole, 47. guide plate, 48. iron sheet, 5. circulation pump, 6. liquid storage tank, 61. miscellaneous pipe, 611. miscellaneous housing, 612. adsorption filter element, 62. turntable, 63. flip plate, 64. driving wheel, 7. conveying pipeline, 8. liquid inlet joint group, 9. liquid outlet joint group, 10. connecting hose, 11. driving motor, 12. U-shaped frame, 13. transmission belt, 14. limit groove, 15. magnet. DETAILED DESCRIPTION

[0032] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model. Example

[0033] like Figures 1 to 8 As shown, a device for screening drugs against colorectal cancer lung metastasis comprises a screening mechanism, wherein a screening component for screening drugs against colorectal cancer lung metastasis is placed on a frame mechanism, and the screening mechanism comprises a liquid phase flow component and a plurality of diversion slots 4;

[0034] The diversion tank 4 includes a bottom tank 41 and a screening plate 42. The screening plate 42 is placed in the bottom tank 41. A plurality of culture recesses 46 are provided on the screening plate 42. A guide plate 47 is connected to the upper plate surface of one end of the screening plate 42. Both ends of the bottom tank 41 are connected to docking ports 45.

[0035] The liquid phase flow component includes a circulation pump 5 and a delivery pipeline 7. The inlet and outlet ends of the circulation pump 5 are connected to the delivery pipeline 7, and the delivery pipeline 7 is also connected to the two ends of the diversion tank 4. A liquid storage tank 6 is arranged on the delivery pipeline 7, and the liquid storage tank 6 is provided with an impurity removal component composed of an adsorption unit, a stirring unit and a power unit. Example

[0036] In addition to all the technical features of the first embodiment, this embodiment also includes:

[0037] The frame structure includes an upper support plate 1, a lower support plate 2 and a column 3. The upper support plate 1 and the lower support plate 2 are parallelly distributed up and down. The column 3 is vertically connected to the four corners of the upper support plate 1 and the lower support plate 2. A diverter trough 4 is placed on the upper support plate 1, and a liquid phase flow component is fixedly connected to the lower support plate 2. The diverter trough 4 and the liquid phase flow component can be arranged in upper and lower layers to facilitate the screening of anti-colorectal cancer lung metastasis drugs by manipulating the diverter trough 4 at the last time.

[0038] A limiting groove 14 is provided at the center of the upper plate surface of the upper support plate 1, and a plurality of magnets 15 are buried in the bottom surface of the limiting groove 14, and an iron sheet 48 is buried in the bottom surface of the bottom groove 41. The corresponding number of diverter grooves 4 required for use can be placed in the limiting groove 14, and the diverter grooves 4 can be fixed in the limiting groove 14 by the magnetic attraction of the magnets 15 and the iron sheet 48 to prevent the diverter grooves 4 from shaking.

[0039] A liquid inlet joint group 8 and a liquid outlet joint group 9 are respectively provided on the short sides of both sides of the upper support plate 1. The bottom ends of the liquid inlet joint group 8 and the liquid outlet joint group 9 are connected to the conveying pipeline 7, and the upper ends of the liquid inlet joint group 8 and the liquid outlet joint group 9 are connected to the docking port 45 through a connecting hose 10, so that the diversion trough 4 and the liquid phase flow component form a closed circulation flow system, and the colorectal cell culture medium as the mobile liquid phase circulates in the system. Example

[0040] In addition to all the technical features of the first embodiment, this embodiment also includes:

[0041] A reserved hole 44 is provided on the bottom surface of the bottom groove 41, and the reserved hole 44 is arranged in a one-to-one correspondence with the culture recessed hole 46. Organoids of normal human lung tissue embedded in Matrigel glue are cultured in the culture recessed hole 46, which can ensure that the screening plate 42 can be placed horizontally on the bottom surface of the bottom groove 41. During the growth process, the lung organoids will release factors specific to lung tissue, which will have a chemotactic effect on the colorectal cancer cells in the flowing liquid phase. The colorectal cancer cells with metastatic ability will move toward the recessed hole, adhere to the surface of Matrigel glue, penetrate the Matrigel glue, enter the glue droplet, interact with the lung organoids, proliferate and form clones. The fluorescence intensity in the glue droplet is detected under a living cell imager to determine the metastatic ability of the metastatic colorectal cancer cells.

[0042] A plurality of guide plates 47 are provided at one end of the screening plate 42 close to the liquid inlet joint group 8 , and the guide plates 47 are arranged in a trumpet shape so that the flowing liquid phase entering the bottom tank 41 can flow evenly on the screening plate 42 under the action of the guide plates 47 . Example

[0043] In addition to all the technical features of the first embodiment, this embodiment also includes:

[0044] The adsorption unit of the impurity removal component includes a collection pipe 61 composed of an impurity collection shell 611 and an adsorption filter element 612. The collection pipe 61 is detachably inserted vertically in the liquid storage tank 6. The collection pipe 61 is tightly attached to the inner wall of the liquid storage tank 6. The adsorption filter element 612 is filled in the collection shell 611. The liquid phase circulating out of the diversion tank 4 flows into the liquid storage tank 6. The impurities contained in the liquid phase can be adsorbed by the adsorption filter element 612, so that the flowing liquid phase entering the diversion tank 4 again can be free of impurities.

[0045] The stirring unit of the impurity removal component includes a turntable 62 and a flip plate 63. The turntable 62 is rotatably connected to the bottom end of the liquid storage tank 6. The flip plate 63 is connected to the turntable 62 in a cross shape, and the upper end of the flip plate 63 is inclined in the direction of rotation. When the turntable 62 rotates, the flip plate 63 can drive the liquid phase in the tank to rotate, and then the impurities in the liquid phase are moved to the vicinity of the tank wall under the action of centrifugal force, so that they can be adsorbed by the adsorption filter element 612 to the maximum extent.

[0046] The power unit of the impurity removal component includes a drive motor 11 and a drive wheel 64. The drive motor 11 is fixedly connected to the U-shaped frame 12 in an inverted state. The U-shaped frame 12 is fixedly welded to the lower support plate 2. The drive wheel 64 is coaxially arranged with the turntable 62 through a connecting shaft, and the rotating shaft of the drive motor 11 and the drive wheel 64 are connected by a transmission belt 13 to provide driving force for the turntable 62, so that the turntable 62 can rotate rapidly in the liquid storage tank 6, driving the liquid phase in the tank to rotate in a vortex shape.

[0047] The side wall of the liquid storage tank 6 is connected to a delivery pipe 7 for inputting the flowing liquid phase, and the upper end of the liquid storage tank 6 is connected to a delivery pipe 7 for outputting the flowing liquid phase, and the delivery pipe 7 for outputting the flowing liquid phase is inserted to the bottom of the liquid storage tank 6. Since the liquid phase in the tank is in a vortex rotation, the liquid phase at the center is in an impurity-free state, and therefore the liquid phase is extracted from the center for circulation.

[0048] Working principle: Fluorescently labeled colorectal cancer cells are in a mobile liquid phase (colorectal cell culture medium), which circulates in a closed system containing a shunt tank. Its flow rate through the shunt tank is close to the flow rate of human venous blood. The shunt tank is provided with a culture concave hole, in which the organoids of normal human lung tissue embedded in Matrigel are cultured. During the growth process, lung organoids will release factors specific to lung tissue, which will have a chemotactic effect on colorectal cancer cells in the mobile liquid phase. Colorectal cancer cells with metastatic ability will move to the concave hole, adhere to the surface of Matrigel, penetrate Matrigel, enter the inside of the gel droplet, interact with the lung organoid, proliferate and form clones. The metastatic ability of metastatic colorectal cancer cells can be determined by detecting the fluorescence intensity in the gel droplet under a live cell imager. When conducting drug screening, the drug (drug group) and control solvent (control group) are added to the mobile liquid phase at appropriate concentrations. If the drug can reduce the anti-apoptosis ability and resistance to liquid phase shear force of colorectal cancer cells in the mobile liquid phase, inhibit the adhesion ability of colorectal cancer cells to the extracellular matrix, inhibit the degradation ability of colorectal cancer cells to the extracellular matrix, and inhibit the colonization ability of colorectal cancer cells in the lung tissue microenvironment, then under a living cell imager, the fluorescence intensity in the drug group droplets will be significantly lower than that in the control group. If the drug has no effect on these links, there will be no significant difference in the fluorescence intensity in the drug group and control group droplets.

[0049] It is obvious to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be regarded as exemplary and non-restrictive from any point of view, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention. Any reference numeral in a claim should not be regarded as limiting the claim to which it relates.

[0050] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A device for screening drugs for preventing colorectal cancer lung metastasis, comprising a screening mechanism, characterized in that: The screening mechanism for screening drugs against colorectal cancer lung metastasis is placed on a frame mechanism, and the screening mechanism includes a liquid phase flow component and a plurality of diversion slots (4); The diversion trough (4) comprises a bottom trough (41) and a screening plate (42), wherein the screening plate (42) is placed in the bottom trough (41), a plurality of culture recesses (46) are provided on the screening plate (42), a guide plate (47) is connected to an upper plate surface at one end of the screening plate (42), and docking ports (45) are connected at both ends of the bottom trough (41); The liquid phase flow component comprises a circulation pump (5) and a delivery pipeline (7); the liquid inlet and outlet ends of the circulation pump (5) are connected to the delivery pipeline (7); and the delivery pipeline (7) is also connected to both ends of the diversion tank (4); a liquid storage tank (6) is provided on the delivery pipeline (7); and the liquid storage tank (6) is provided with an impurity removal component consisting of an adsorption unit, an agitation unit and a power unit.

2. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 1, characterized in that: The frame structure comprises an upper support plate (1), a lower support plate (2) and a column (3); the upper support plate (1) and the lower support plate (2) are arranged in parallel up and down; the column (3) is vertically connected to the four corners of the upper support plate (1) and the lower support plate (2); a diversion trough (4) is arranged on the upper support plate (1); and a liquid phase flow component is fixedly connected to the lower support plate (2).

3. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 2, characterized in that: A limiting groove (14) is provided at the center of the upper plate surface of the upper support plate (1), a plurality of magnets (15) are embedded in the bottom surface of the limiting groove (14), and an iron sheet (48) is embedded in the bottom surface of the bottom groove (41).

4. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 2, characterized in that: A liquid inlet joint group (8) and a liquid outlet joint group (9) are respectively provided on the short sides of both sides of the upper support plate (1); the bottom ends of the liquid inlet joint group (8) and the liquid outlet joint group (9) are both connected to the delivery pipeline (7); and the upper ends of the liquid inlet joint group (8) and the liquid outlet joint group (9) are both connected to the docking port (45) via a connecting hose (10).

5. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 1, characterized in that: The bottom surface of the bottom groove (41) is provided with reserved holes (44), and the reserved holes (44) are arranged in a one-to-one correspondence with the culture recessed holes (46), and the organoids of normal human lung tissue embedded in Matrigel are cultured in the culture recessed holes (46).

6. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 5, characterized in that: A plurality of guide plates (47) are provided at one end of the screening plate (42) close to the liquid inlet joint group (8), and the guide plates (47) are arranged in a trumpet shape.

7. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 1, characterized in that: The adsorption unit of the impurity removal component comprises an impurity collecting pipe (61) composed of an impurity collecting shell (611) and an adsorption filter element (612); the impurity collecting pipe (61) is detachably vertically inserted in the liquid storage tank (6); the impurity collecting pipe (61) is closely attached to the inner wall of the liquid storage tank (6); and the adsorption filter element (612) is filled in the impurity collecting shell (611).

8. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 7, characterized in that: The stirring unit of the impurity removal component comprises a rotating disk (62) and a flip plate (63); the rotating disk (62) is rotatably connected to the bottom end of the liquid storage tank (6); the flip plate (63) is connected to the rotating disk (62) in a cross shape, and the upper end of the flip plate (63) is inclined in the direction of rotation.

9. The device for screening drugs for preventing colorectal cancer lung metastasis according to claim 7, characterized in that: The power unit of the impurity removal component comprises a drive motor (11) and a drive wheel (64); the drive motor (11) is fixedly connected to a U-shaped frame (12) in an inverted state; the U-shaped frame (12) is fixedly welded to a lower support plate (2); the drive wheel (64) is coaxially arranged with a turntable (62) via a connecting shaft; and the rotating shaft of the drive motor (11) and the drive wheel (64) are connected in transmission via a transmission belt (13).

10. The device for screening drugs against colorectal cancer lung metastasis according to claim 1, characterized in that: The side wall of the liquid storage tank (6) is connected to a delivery pipe (7) for inputting a mobile liquid phase, the upper end of the liquid storage tank (6) is connected to a delivery pipe (7) for outputting a mobile liquid phase, and the delivery pipe (7) for outputting a mobile liquid phase is inserted into the bottom of the liquid storage tank (6).

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