Mouse Weber's gland duct source oropharyngeal cancer model and construction method thereof

Abstract

The invention discloses a mouse Weber's gland duct source oropharyngeal cancer model and a construction method thereof. The construction method comprises the following steps: selecting a male BALB / c mouse of 6 weeks old; placing the 4NQO aqueous solution in a light-proof bottle for free drinking of the mouse; from the 17th week, putting the mixed solution of 4NQO and glucose into a light-proof bottle for free drinking of the mouse; after the twentieth week, a glucose aqueous solution is adopted for free drinking of the mice, and the mice are fed with raw peanuts; after the mouse is raised for 27 weeks, the mouse Weber's gland duct source oropharyngeal cancer model is obtained.According to the construction method, through limitation of all the steps, construction of the tumor at the Weber's gland duct part of the tongue root of the mouse can be achieved, the construction method is easy to operate and high in success rate, the survival rate of the mouse in the later period is increased, and a new research direction is developed for research of oropharyngeal cancer in the future.

Description

Technical Field

[0001] The present invention relates to the technical field of animal models, and particularly relates to an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice and a method for constructing the same. Background Art

[0002] Oropharyngeal squamous cell carcinoma mainly originates from the oral pharyngeal mucosal epithelium such as the root of the tongue, palatine tonsil, posterior pharyngeal wall, and soft palate, especially the palatine tonsil and the root of the tongue are the most common. Its incidence is closely related to chemical stimuli such as long-term smoking and alcoholism, as well as human papilloma virus (HPV) infection. At present, a major problem faced in the research related to oropharyngeal cancer is the lack of relevant animal models. As an important means for studying the pathogenesis of diseases and exploring treatment methods, the absence of animal models greatly limits the in-depth study of oropharyngeal cancer.

[0003] Weber's gland, as a pure mucous gland located at the root of the tongue, has been found to be an important origin of oropharyngeal cancer in research. This discovery can provide new ideas for the clinical treatment of oropharyngeal cancer; in addition, the glandular duct of Weber's gland in the human root of the tongue is found to have an opening pattern and histological structure similar to that of mice, which provides a solid anatomical and histological basis for using mice to study the glandular duct of Weber's gland; however, due to the lack of relevant animal models, subsequent related research is difficult to carry out.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to provide an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice and a method for constructing the same. The construction method of the present invention is simple to operate and has a high success rate, and can provide a powerful tool for studying oropharyngeal cancer related to the glandular duct of Weber's gland at the root of the tongue.

[0006] In order to achieve the above object of the present invention, the following technical solutions are specifically adopted:

[0007] The first aspect of the present invention provides a method for constructing an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice, and the method for constructing the oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice includes the following steps:

[0008] (a) Select male BALB / c mice at 6 weeks of age;

[0009] (b) Place an aqueous solution of 4NQO (4-nitroquinoline-N-oxide) in a light-proof bottle for the mice to drink freely;

[0010] (c) Starting from the 17th week, place a mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely;

[0011] (d) After the 20th week, a glucose aqueous solution was provided for the mice to drink freely, and raw peanuts were fed.

[0012] (e) When the mice were raised until 27 weeks, an oropharyngeal cancer model derived from the glandular ducts of Weber's gland in mice was obtained.

[0013] Preferably, in the step (b), the concentration of the 4NQO aqueous solution is 200 mg / L.

[0014] Preferably, in the step (c), the concentration of 4NQO in the mixed solution of 4NQO and glucose is 200 mg / L, and the concentration of glucose is 5%.

[0015] Preferably, in the step (d), the concentration of the glucose aqueous solution is 5%.

[0016] Preferably, in the step (c), the feeding method of the raw peanuts is unlimited.

[0017] Preferably, the success rate of constructing the oropharyngeal cancer model derived from the glandular ducts of Weber's gland in mice can reach 100%.

[0018] The second aspect of the present invention provides an oropharyngeal cancer model derived from the glandular ducts of Weber's gland in mice constructed by the above construction method.

[0019] Compared with the prior art, the beneficial effects of the present invention at least include:

[0020] Through the limitation of each step, the construction method of the present invention can realize the construction of tumors in the glandular ducts of Weber's gland in the root of the tongue of mice. Moreover, the construction method of the present invention is simple in operation, has a high success rate, and improves the survival rate of mice in the later stage, opening up a new research direction for the future research of oropharyngeal cancer. Description of the Drawings

[0021] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for use in the description of the specific embodiments or the prior art. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn to actual scale.

[0022] Figure 1 It is a tissue section diagram (200×) of each stage of the occurrence and development of squamous cell carcinoma derived from the glandular ducts of Weber's gland in the root of the tongue of mice in the experimental example of the present invention;

[0023] Figure 2 It is a picture of the root of the tongue of the mouse in Example 1 of the experimental example of the present invention 27 weeks after modeling;

[0024] Figure 3In the experimental example of the present invention, the situation of the human lingual root Weber's gland (traditional mode) opening into the crypts of the lingual tonsil, where the glandular duct of the Weber's gland opens into the crypts of the lingual tonsil and the epithelial cells of its glandular duct (red arrow) are continuous with the unique reticular epithelium of the tonsil (green arrow);

[0025] Figure 4 This is a schematic diagram of the opening mode of the glandular duct of the Weber's gland in the human lingual root directly opening into the squamous epithelium of the dorsal tongue between the lingual tonsils in the experimental example of the present invention. The epithelial cells of the glandular duct are directly continuous with the squamous epithelium of the dorsal tongue (40×);

[0026] Figure 5 This is the Weber's gland of a mouse in the experimental example of the present invention. In rodents, the Weber's gland directly opens into the dorsal epithelium of the lingual root. The epithelial cells of the glandular duct are directly continuous with the squamous epithelium of the dorsal tongue (200×). Detailed implementation manners

[0027] Next, the embodiments of the technical solutions of the present invention will be described in detail in conjunction with the embodiments. The following embodiments are only used to illustrate the technical solutions of the present invention more clearly, so they are only examples and cannot be used to limit the protection scope of the present invention.

[0028] It should be noted that unless otherwise specified, the technical terms or scientific terms used in this application should have the ordinary meanings understood by those skilled in the art to which the present invention belongs.

[0029] The embodiment of the present invention provides a method for constructing an oropharyngeal cancer model derived from the glandular duct of the Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular duct of the Weber's gland in mice includes the following steps:

[0030] (a) Select male BALB / c mice at 6 weeks of age;

[0031] (b) Place the 4NQO aqueous solution in a light-proof bottle for the mice to drink freely;

[0032] (c) Starting from the 17th week, place the mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely;

[0033] (d) After the 20th week, use the glucose aqueous solution for the mice to drink freely and feed raw peanuts;

[0034] (e) Wait until the mice are raised to 27 weeks to obtain an oropharyngeal cancer model derived from the glandular duct of the Weber's gland in mice.

[0035] The construction method of the present invention can realize the construction of tumors in the glandular ducts of Weber's glands at the root of the mouse tongue through the limitations of each step. Moreover, the construction method of the present invention is simple to operate, has a high success rate, and improves the survival rate of mice in the later stage, opening up a new research direction for the future research of oropharyngeal cancer.

[0036] In one embodiment, in the step (b), the concentration of the 4NQO aqueous solution can be 200 mg / L.

[0037] In one embodiment, in the step (c), the concentration of 4NQO in the mixed solution of 4NQO and glucose can be 200 mg / L, and the concentration of glucose can be 5%.

[0038] In one embodiment, in the step (d), the concentration of the glucose aqueous solution can be 5%.

[0039] In one embodiment, in the step (c), the feeding method of the raw peanuts is not limited in quantity.

[0040] In one embodiment, the success rate of constructing the oropharyngeal cancer model derived from the glandular ducts of Weber's glands in mice can reach 100%.

[0041] Another embodiment of the present invention provides an oropharyngeal cancer model derived from the glandular ducts of Weber's glands in mice constructed by the above construction method.

[0042] The technical solution of the present invention will be further described in detail below through specific examples.

[0043] Example 1

[0044] This example is a construction method of an oropharyngeal cancer model derived from the glandular ducts of Weber's glands in mice. The construction method of the oropharyngeal cancer model derived from the glandular ducts of Weber's glands in mice includes the following steps:

[0045] (a) Select 6-week-old male BALB / c mice;

[0046] (b) Place the 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0047] (c) Starting from the 17th week, place the mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely, wherein the concentration of 4NQO in the mixed solution of 4NQO and glucose is 200 mg / L, and the concentration of glucose is 5%;

[0048] (d) After the 20th week, use the glucose aqueous solution with a concentration of 5% for the mice to drink freely, and feed raw peanuts, wherein the feeding method of the raw peanuts is not limited in quantity;

[0049] (e) Feed until 27 weeks to obtain an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice.

[0050] Comparative Example 1

[0051] This comparative example is a method for constructing an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice includes the following steps:

[0052] (a) Select 6-week-old male BALB / c mice;

[0053] (b) Place a 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0054] (c) Starting from the 17th week, place the 4NQO aqueous solution in a light-proof bottle for the mice to drink freely, wherein the concentration of 4NQO in the 4NQO aqueous solution is 200 mg / L;

[0055] (d) After the 20th week, use drinking water for the mice to drink freely and feed raw peanuts, and the feeding method of the raw peanuts is not limited in quantity;

[0056] (e) Feed until 27 weeks to obtain an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice.

[0057] Comparative Example 2

[0058] This comparative example is a method for constructing an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice includes the following steps:

[0059] (a) Select 6-week-old male BALB / c mice;

[0060] (b) Place a 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0061] (c) Starting from the 17th week, place a mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely, wherein the concentration of 4NQO in the mixed solution of 4NQO and glucose is 200 mg / L, and the concentration of glucose is 5%;

[0062] (d) After the 20th week, use a 5% glucose aqueous solution for the mice to drink freely;

[0063] (e) Feed until 27 weeks to obtain an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice.

[0064] Comparative Example 3

[0065] This comparative example is a method for constructing an oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice includes the following steps:

[0066] (a) Select male BALB / c mice at 6 weeks of age;

[0067] (b) Place a 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0068] (c) After the 20th week, use drinking water for the mice to drink freely;

[0069] (d) Wait until the mice are raised to 27 weeks to obtain an oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice.

[0070] Comparative Example 4

[0071] This comparative example is a method for constructing an oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice includes the following steps:

[0072] (a) Select male BALB / c mice at 6 weeks of age;

[0073] (b) Place a 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0074] (c) Starting from the 17th week, place a mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely. Among them, the concentration of 4NQO in the mixed solution of 4NQO and glucose is 200 mg / L, and the concentration of glucose is 10%;

[0075] (d) After the 20th week, use a 10% glucose aqueous solution for the mice to drink freely and feed raw peanuts. Among them, the feeding method of raw peanuts is unlimited;

[0076] (e) Wait until the mice are raised to 27 weeks to obtain an oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice.

[0077] Comparative Example 5

[0078] This comparative example is a method for constructing an oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice. The method for constructing the oropharyngeal cancer model derived from the glandular ducts of the Weber's gland in mice includes the following steps:

[0079] (a) Select male BALB / c mice at 6 weeks of age;

[0080] (b) Place a 4NQO aqueous solution with a concentration of 200 mg / L in a light-proof bottle for the mice to drink freely;

[0081] (c) Starting from the 17th week, a mixed solution of 4NQO and glucose was placed in a light-proof bottle for the mice to drink freely. Among them, the concentration of 4NQO in the mixed solution of 4NQO and glucose was 200 mg / L, and the concentration of glucose was 5%.

[0082] (d) After the 20th week, an aqueous glucose solution with a concentration of 5% was provided for the mice to drink freely, and raw melon seeds were fed. Among them, the feeding method of the raw melon seeds was not limited in quantity.

[0083] (e) When the mice were raised to 27 weeks, an oropharyngeal cancer model of mouse Weber's gland duct origin was obtained.

[0084] Experimental Example

[0085] Three hundred and fifty 6-week-old male BALB / c mice were selected and randomly divided into 7 groups.

[0086] Among them, 6 groups of mice were respectively used to construct the oropharyngeal cancer model of mouse Weber's gland duct origin by the methods of Example 1 and Comparative Examples 1-5; the other group was used as a blank control.

[0087] After 27 weeks and 30 weeks, the survival rate of the mice and the success rate of the oropharyngeal cancer model of mouse Weber's gland duct origin were counted.

[0088] Five mice were randomly selected and sacrificed, and then verified by histology to see if canceration from Weber's gland duct generally occurred, and the construction success rate was calculated.

[0089] (200×) Histological section diagrams of various stages of the occurrence and development of squamous cell carcinoma originating from Weber's gland duct in the mouse tongue root are as Figure 1 shown, Figure 1 wherein, A is normal, B is abnormal hyperplasia, and C is squamous cell carcinoma.

[0090] The picture of the tongue root of the mouse after modeling for 27 weeks in Example 1 is as Figure 2 shown;

[0091] The situation where the Weber's gland (traditional mode) in the human tongue root opening in the crypt of the lingual tonsil, and the duct epithelium (red arrow) of the Weber's gland duct continues with the specific reticular epithelium (green arrow) of the tonsil is as Figure 3 shown;

[0092] (40×) The opening mode where the duct of the Weber's gland in the human tongue root directly opens into the lingual dorsal squamous epithelium between the lingual tonsils, and the duct epithelium directly continues with the squamous epithelium of the lingual dorsum (not previously recorded) is as Figure 4 described, Figure 4In it, A and B are the same tissue. However, the whole picture cannot be captured under a 40 - fold microscope, so two pictures are put together to prove that this gland is the small mucous gland in the root of the tongue - Weber's gland. Figure A shows that the human gland duct directly opens into the dorsal mucosa of the root of the tongue, and Figure B shows the lingual tonsil near it.

[0093] (200×) The Weber's gland of mice. In rodents, the Weber's gland directly opens into the dorsal epithelium of the root of the tongue, and the epithelium of the gland duct is directly continuous with the squamous epithelium of the dorsal tongue (as Figure 5 shown).

[0094] It can be seen from Figures 3 to 5 that:

[0095] There is a new opening pattern in the gland duct of the Weber's gland in the root of the human tongue, which is similar to that of mice, directly opening into the dorsal epithelium of the root of the tongue. This provides a solid histological basis for selecting mice to construct an oropharyngeal cancer model derived from the gland duct of the Weber's gland.

[0096] The statistical results are shown in Table 1:

[0097] Table 1

[0098]

[0099]

[0100] It can be seen from Table 1 that:

[0101] The method for constructing an oropharyngeal cancer model derived from the gland duct of the Weber's gland of mice in this application can successfully construct an oropharyngeal cancer model derived from the gland duct of the Weber's gland of mice, and the survival rate and success rate of the mice are both significantly improved.

[0102] Finally, it should be noted that: the above - mentioned embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and the description of the present invention.

Claims

1. A method for constructing an oropharyngeal cancer model derived from the glandular duct of Weber's gland in mice, characterized in that, It includes the following steps: (a) Select male BALB / c mice at 6 weeks old; (b) Place the 4NQO aqueous solution in a light-proof bottle for the mice to drink freely; (c) Starting from the 17th week, place the mixed solution of 4NQO and glucose in a light-proof bottle for the mice to drink freely; (d) After the 20th week, use the glucose aqueous solution for the mice to drink freely and feed raw peanuts; (e) Wait until the mice are raised to 27 weeks to obtain an oropharyngeal cancer model derived from the glandular duct of Weber's gland of the mice.

2. The method for constructing a mouse oropharyngeal cancer model derived from the glandular ducts of Weber's glands according to claim 1, wherein In the step (b), the concentration of the 4NQO aqueous solution is 200 mg / L.

3. The method for constructing a mouse oropharyngeal cancer model derived from the glandular ducts of Weber's glands according to claim 1, characterized in that In the step (c), the concentration of 4NQO in the mixed solution of 4NQO and glucose is 200 mg / L, and the concentration of glucose is 5%.

4. The method for constructing a mouse oropharyngeal cancer model derived from the glandular ducts of Weber's gland according to claim 1, wherein In the step (d), the concentration of the glucose aqueous solution is 5%.

5. The method for constructing a mouse oropharyngeal cancer model derived from the glandular ducts of Weber's glands according to claim 1, characterized in that, In the step (c), the feeding method of the raw peanuts is unlimited.

6. The method for constructing a mouse oropharyngeal cancer model derived from the glandular ducts of Weber's glands according to claim 1, wherein, The success rate of constructing the oropharyngeal cancer model derived from the glandular duct of Weber's gland of the mice can reach 100%.

7. An oropharyngeal cancer model derived from the glandular duct of Weber's gland of the mice constructed by the construction method according to claims 1 to 6.

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