论文部分内容阅读
目的探讨重组质粒p DC316h IL-24基因联合顺铂通过m TOR/P70S6K信号通路抑制人宫颈癌siha细胞淋巴转移。方法将造模成功的人宫颈癌siha细胞雌性裸鼠模型随机分为6组,每组7只:PBS对照组、p DC316h IL-24组、低剂量顺铂组、高剂量顺铂组、p DC316h IL-24+低剂量顺铂组、p DC316h IL-24+高剂量顺铂组。观察裸鼠一般状况及淋巴结转移情况,实验结束后采用聚合酶链反应(PCR)法测定移植瘤中IL-24 m RNA的表达,CK角蛋白检测获取的淋巴组织有否癌浸润,免疫荧光法检测转移淋巴组织中m TOR和P70S6K蛋白表达,并用Western blot印迹法对其表达做定量分析。结果 IL-24基因成功转染及表达;CK角蛋白检测获取的各组淋巴组织有癌浸润;干预后各组淋巴转移数有差异,其中联合组淋巴结转移数明显低于对照组(P<0.05),而IL-24+低剂量顺铂组、IL-24+高剂量顺铂组淋巴转移数无统计学差异(P>0.05);免疫荧光法成功检测到各组转移淋巴组织中m TOR/P70S6K的表达,采用Western印迹法进行定量分析,与其他组比较,联合组表达显著下降,差异有统计学意义(P<0.05),且m TOR与P70S6K表达呈正相关性(r=0.968,P<0.001)。结论 IL-24+低剂量顺铂治疗能达到IL-24+高剂量顺铂的治疗效果;IL-24+低剂量顺铂能明显抑制宫颈癌siha细胞淋巴转移;IL-24联合顺铂在体内通过抑制m TOR/P70S6K信号通路的活性促进细胞凋亡,从而抑制人宫颈癌siha细胞的淋巴转移。
Objective To investigate the inhibitory effect of recombinant plasmid p DC316h IL-24 gene combined with cisplatin on lymphatic metastasis of human cervical carcinoma siha cells through the mTOR / P70S6K signaling pathway. Methods The successful model of human cervical cancer siha cells in nude mice were randomly divided into 6 groups with 7 mice in each group: PBS control group, p DC316h IL-24 group, low-dose cisplatin group, high-dose cisplatin group, p DC316h IL-24 + low-dose cisplatin group, p DC316h IL-24 + high-dose cisplatin group. The general condition and lymph node metastasis of nude mice were observed. After the experiment, the expression of IL-24 m RNA in the transplanted tumor was detected by polymerase chain reaction (PCR), and the lymphoid tissue infiltration was detected by CK keratinocytes. Immunofluorescence The expressions of m TOR and P70S6K in metastatic lymphoid tissue were detected by Western blotting and the expression was quantitatively analyzed. Results The IL-24 gene was successfully transfected and expressed. The lymphoid tissues in each group obtained by CK keratin detection had a different degree of lymph node metastasis. The number of lymph node metastasis in the combined group was significantly lower than that in the control group (P <0.05 ), While there was no significant difference in IL-24 + low-dose cisplatin group and IL-24 + high-dose cisplatin group (P> 0.05). Immunofluorescence method was used to detect m TOR / The expression of P70S6K was quantitatively analyzed by Western blot. Compared with other groups, the expression of P70S6K in the combination group was significantly decreased (P <0.05), and the correlation between mTOR and P70S6K expression was significant (r = 0.968, P < 0.001). Conclusion IL-24 + low-dose cisplatin treatment can achieve the effect of IL-24 + high-dose cisplatin treatment; IL-24 + low-dose cisplatin can significantly inhibit cervical cancer siha cell lymphatic metastasis; IL-24 combined with cisplatin in vivo By inhibiting the activity of the mTOR / P70S6K signaling pathway, apoptosis is inhibited, thereby inhibiting lymphatic metastasis in human cervical cancer siha cells.