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目的:探讨膜联蛋白a3(ANXA3)在结直肠癌HT-29细胞株和HT-29奥沙利铂(L-OHP)耐药细胞株中的表达。方法:采用药物浓度梯度递增间歇诱导法建立人结直肠癌耐L-OHP的耐药细胞株HT-29/LOHP;CCK-8测定L-OHP对HT-29/L-OHP和HT-29细胞株的半数抑制率(IC50);RT-PCR分别检测HT-29细胞和HT-29/L-OHP耐药细胞中ANXA3 RNA的表达水平;蛋白质印迹法检测ANXA3在HT-29细胞和HT-29/L-OHP细胞中的表达差异。结果:(1)成功建立了耐药细胞株HT-29/L-OHP,其耐药指数为3.72;(2)HT-29/L-OHP细胞株的ANXA3 mRNA表达水平(0.897±0.260)高于HT-29细胞株(0.513±0.203)(P<0.05);(3)耐药细胞株HT-29/L-OHP的ANXA3蛋白表达水平高于亲本细胞株HT-29。结论:ANXA3在耐药细胞株中RNA基因水平和蛋白水平表达显著高于亲本细胞株HT-29。
Objective: To investigate the expression of annexin a3 (ANXA3) in colorectal cancer HT-29 cells and HT-29 oxaliplatin (L-OHP) resistant cell lines. Methods: The HT-29 / LOHP resistant human colorectal cancer cell line L-OHP was established by intermittent induction with drug concentration gradient. The effects of L-OHP on HT-29 / L-OHP and HT-29 cells (IC50). The expression of ANXA3 RNA in HT-29 cells and HT-29 / L-OHP-resistant cells was detected by RT-PCR. Western blotting was used to detect the expression of ANXA3 in HT-29 cells and HT- 29 cells / L-OHP cells. Results: (1) The resistant cell line HT-29 / L-OHP was successfully established and the drug resistance index was 3.72. (2) The expression of ANXA3 mRNA in HT-29 / L-OHP cell line (0.897 ± 0.260) (0.513 ± 0.203) (P <0.05). (3) The expression level of ANXA3 in HT-29 / L-OHP group was higher than that in parental cell line HT-29. CONCLUSION: The expression of ANXA3 in the drug-resistant cell line was significantly higher than that of the parent cell line HT-29.