Survivin反义核酸联合紫杉醇对皮下荷瘤Balb/c小鼠模型的治疗作用
The therapeutic effects of survivin antisense nucleic acid combined with paclitaxel on subcutaneous xenograft mouse model of Balb/c

摘要：目的 研究survivin反义核酸与紫杉醇联合应用对皮下荷瘤Bal b/c小鼠模型的治疗作用，并初步探讨对其抗癌作用的机制。方法 在Bal b/c小鼠皮下注射C26细胞，建立皮下瘤模型，采用瘤内注射的方式，将实验分空白组(C)、lipo2000对照组(L)、紫杉醇组(T)、survivin反义核酸组(A)、survivin反义核酸联合紫杉醇组(A+T)5个不同组，观察肿瘤的生长状态，TUNEL法检测凋亡细胞，Western blot法检测survivin蛋白表达。结果 ①各治疗组均达到了(T/C)<60%，与L组比较差异有统计学意义(P<0.05)，体内证实给药干预有效；小鼠瘤重的抑瘤率结果显示，与C组比较，T、A、A+T各给药组均能抑制小鼠瘤重，差异具有统计学意义(P<0.05)，从抑制肿瘤质量增长方面而言，二者联合用药[瘤重抑瘤率为(54.16±0.32)%]将紫杉醇[瘤重抑瘤率为(21.82±0.84)%]的抗癌活性提高了59%以上；②TUNEL法检测凋亡细胞：空白对照组几乎没有肿瘤细胞凋亡。T组及A组有一定量的凋亡细胞，以上试验结果提示，紫杉醇具有促进肿瘤细胞凋亡的能力，A+T不仅加强了对肿瘤细胞的杀伤作用，而且二者的协同作用可能对肿瘤耐药性有所影响，最终使得其促肿瘤细胞凋亡的作用尤为显著；③survivin蛋白表达：结果显示，A+T组survivin蛋白的表达明显降低，而不影响β-actin的表达，C组和L组相比无明显变化，T组、A组、A+T组A值的比值分别为0.895±0.011、0.704±0.121、0.345±0.019，经方差分析，A+T组与C、L、A、T组的表达量差异具有统计学意义(P<0.05)。结论 survivin反义核酸与紫杉醇联合用药，可能通过下调survivin蛋白的表达从而促进肿瘤细胞凋亡，联合用药可降低机体耐药性，发挥协同作用。
ABSTRACT: Objective To explore the therapeutic effects of combined application of survivin antisense nucleic acid and taxol in subcutaneous xenograft mouse model of Balb/c and to preliminarily investigate the mechanism of the anticancer effects. Methods The model of subcutaneous tumor was established by hypodermic injection of C26 cells into Bal b/c mice. The mice were then randomly divided into five groups through the internal tumor injection: the blank group (C), lipo2000 group (L), paclitaxel group (T), survivin antisense nucleic acid group (A), and survivin antisense nucleic acid combined with paclitaxel group (A+T). We observed tumor growth, determined cell apoptosis by TUNEL method, and detected the expression of survivin by Western blot. Results ① All treatment groups had T/C<60%, which was significantly different from that of group L (P<0.05); the intervention was proved effective in vivo. The tumor inhibition rate of mice tumor weight showed that there were significantly curative effects in groups T, A and A+T compared with that in group C (P<0.05). The antitumor activity of paclitaxel (tumor inhibition rate of 21.82%±0.84%) could be improved by more than 59% through combination therapy (tumor inhibition rate of 54.16%±0.32%) concerning inhibition of tumor weight growth. ② TUNEL method detected apoptotic cells: The tumor cells hardly had apoptosis in the blank group while T group and A group had a certain number of apoptotic cells. The experiment results suggested that PTX could promote tumor cell apoptosis, and that not only A+T strengthened the effect in killing tumor cells, but also the synergy of both could influence tumor resistance and ultimately make the effect in promoting tumor cell apoptosis conspicuous. ③ The expression