论文部分内容阅读
目的:体外观察羟基喜树碱(HCPT)联合奥沙利铂(L-OHP)对人大肠癌耐药细胞株LoVo/5-FU增殖抑制及诱导凋亡作用,并初步探讨其诱导凋亡方面机制。方法:MTT法测定HCPT联合L-OHP对LoVo/5-FU细胞增殖抑制作用,并根据中效原理判断两药合用时的效应。流式细胞仪观察两药对LoVo/5-FU细胞的凋亡诱导作用。半定量RT-PCR方法检测单药及联合作用后X染色体连锁的凋亡抑制蛋白(XIAP)mRNA表达变化。结果:HCPT联合L-OHP对LoVo/5-FU细胞有明显的抑制增殖作用,细胞增殖的抑制率与药物浓度呈正相关。在抑制率(fa)>0.18时合用指数(CI)<1,两药表现为协同作用。HCPT浓度分别为0.02、0.04和0.08 mg/L。作用48 h后,LoVo/5-FU细胞凋亡率分别为5.92±0.58、9.93±0.62和3.76±0.45。当0.04 mg/L HCPT联合L-OHP作用48 h后,其凋亡率为18.44±0.57,联合组与单药组比较差异有统计学意义,P<0.05。耐药细胞LoVo/5-FU较亲本细胞LoVo高表达XIAP mRNA(0.88±0.03vs0.11±0.02,P<0.01),两药联合作用后XIAP mRNA降低明显为0.21±0.03,P<0.01。结论:HCPT联合L-OHP能够协同抑制LoVo/5-FU细胞增殖,其机制可能与诱导细胞凋亡有关,抑制XIAP表达发挥作用。
OBJECTIVE: To observe the effects of HCPT and L-OHP on the proliferation and apoptosis of human colorectal cancer cell line LoVo / 5-FU in vitro and to investigate the role of HCPT and oxaliplatin in inducing apoptosis mechanism. Methods: The inhibitory effect of HCPT combined with L-OHP on the proliferation of LoVo / 5-FU cells was determined by MTT assay. Flow cytometry was used to observe the apoptosis-inducing effect of the two drugs on LoVo / 5-FU cells. Semiquantitative RT-PCR was used to detect the change of X-linked inhibitor of apoptosis protein (XIAP) mRNA expression after monotherapy and combined treatment. Results: HCPT combined with L-OHP significantly inhibited the proliferation of LoVo / 5-FU cells. The inhibition rate of cell proliferation was positively correlated with drug concentration. The combination index (CI) <1 at the inhibition rate (fa)> 0.18 showed synergy between the two drugs. HCPT concentrations were 0.02, 0.04 and 0.08 mg / L, respectively. After 48 h treatment, the apoptosis rates of LoVo / 5-FU cells were 5.92 ± 0.58, 9.93 ± 0.62 and 3.76 ± 0.45, respectively. When the concentration of 0.04 mg / L HCPT combined with L-OHP for 48 h, the apoptosis rate was 18.44 ± 0.57, the difference between the combined group and the single drug group was statistically significant, P <0.05. Compared with LoVo, the expression of XIAP mRNA in LoVo / 5-FU cells (0.88 ± 0.03vs0.11 ± 0.02, P <0.01) was significantly higher than that in LoVo cells. The decrease of XIAP mRNA was 0.21 ± 0.03 (P <0.01). CONCLUSION: HCPT combined with L-OHP can synergistically inhibit the proliferation of LoVo / 5-FU cells. The mechanism may be related to the induction of apoptosis and the inhibition of XIAP expression.