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

Inactive Publication Date: 2015-10-21
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although targeted drugs can significantly reduce tumor volume and prolong the median progression-free survival and overall survival of patients with advanced renal cell carcinoma, they cannot maintain long-lasting efficacy, and most patients will develop drug resistance or even progress within 5 to 10 months of treatment. Therefore, it is

Method used

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  • Renal cancer sunitinib drug-resistant cell system and establishing method thereof
  • Renal cancer sunitinib drug-resistant cell system and establishing method thereof
  • Renal cancer sunitinib drug-resistant cell system and establishing method thereof

Examples

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Effect test

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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Abstract

The invention relates to the technical field of biomedicine, and in particular to a renal cancer sunitinib drug-resistant cell system and an establishing method and application of the renal cancer sunitinib drug-resistant cell system. 786-O cells and ACHN cells of a human renal clear cell carcinoma system are adopted as objects for establishing a subcutaneous tumor-bearing nude mouse model; the clinical conventional dose of the sunitinib is used for treating a tumor-bearing nude mouse; and through multiple times of in-vivo continuous passage and drug pressure screening, the renal cancer drug-resistant cell system free of restraints of the sunitinib in vitro and vivo is finally obtained through separate culture and named as 7SuR and ACSuR. The established renal cancer sunitinib drug-resistant cell system can better simulate clinical drug resistance and provides an important platform for studying renal cancer targeted drug resistance mechanism, reversing renal cancer cell drug resistance and developing and evaluating new anti-cancer drugs.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a sunitinib drug-resistant cell line of renal cancer and its construction method and application. Background technique [0002] Kidney cancer accounts for about 3% of adult malignant tumors, and its incidence rate ranks second among urological tumors in China. Its main pathological type is clear cell carcinoma, about 30% of patients are found to have distant metastasis at the time of diagnosis, and the incidence of postoperative distant metastasis is as high as 30% to 40%. Because renal cancer is not sensitive to traditional radiotherapy and chemotherapy, cytokine therapy based on interleukin-2 and interferon-α once became the main treatment for advanced renal cancer, but it only works in less than 20% of patients. effective, with significant side effects. [0003] In recent years, the small molecule targeted drugs represented by sunitinib have greatly improved the treatmen...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 曲乐王林辉刘冰陈程吴震杰张雯
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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