Renal cancer sunitinib drug-resistant cell system and establishing method thereof
A kidney cancer sunitinib and drug-resistant cell technology, applied in the field of biomedicine, can solve the problems of inability to maintain lasting efficacy, lack, and reduce tumor volume in advanced renal cell carcinoma patients, such as progression and survival, and achieve high scientific research and production applications. effect of value
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Embodiment 1
[0037] Example 1: Establishment of human renal carcinoma cell line 786-O and ACHN nude mouse tumor-bearing models
[0038] 1) After resuscitating human renal carcinoma cell line 786-O and ACHN cells, use DMEM culture medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Cultivate in an incubator, change the medium every 2-3 days, and subculture and expand the culture when the cells grow to a density of about 80%.
[0039] 2), collect the human kidney cancer cell line 786-O and ACHN cells in the logarithmic growth phase at 3×10 6 Each spot was inoculated subcutaneously on both sides of the scapula of 8-week-old nude mice, in which 786-O was inoculated subcutaneously on the left scapula, and ACHN was inoculated subcutaneously on the right scapula.
Embodiment 2
[0040] Example 2: Sunitinib treatment and serial passage in vivo of human renal carcinoma cell line 786-O and ACHN tumor-bearing nude mice
[0041] 1) The volume of transplanted tumor in nude mice reaches 200mm 3 Afterwards, the nude mice were randomly divided into two groups, with 5 tumor-bearing nude mice in each group. The sunitinib group was given intragastric administration of 40 mg / kg sunitinib every day, and the control group was given intragastric administration of equal volume of saline every day for 4 weeks. medicine for 2 weeks. At the same time, the long and short diameters of the implanted tumors were measured with a vernier caliper at the designated time points.
[0042] 2) The tumor-bearing mice were killed by cervical dislocation, and the nude mice were sterilized with alcohol cotton balls, especially the tumor inoculation site. The skin of the nude mice was cut with ophthalmic scissors, and the tumor tissue was aseptically stripped and placed in pre-cooled sa...
Embodiment 3
[0048] Embodiment 3: the cytomorphological observation of contrast and drug-resistant cell
[0049] Observation of live cell morphology with an inverted phase-contrast microscope: 786-O and ACHN control cells in the logarithmic growth phase and 7SuR and ACSuR drug-resistant cells were taken respectively, and the live cell morphology was observed and photographed under an inverted microscope (×200 times) after changing the medium. Compared with 786-O, 7SuR showed obvious spindle-shaped changes and increased in size through light microscopy; ACHN cells were epithelioid, with regular size and shape, while ACSuR cells were slightly larger, irregular in shape, and polygonal with long pseudopods. Enlarged size with giant cells (eg, figure 2 shown).
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