【摘要】 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。更多还原

【Abstract】 Objective To investigate the inhibition of IL-24 in combination with cisplatin on human uterine lymph node metastasis. Methods Nude mice xenograft models were established with human cervical cancer siha cells, and then animal models were randomly divided into 6 groups with 7 mice in each group:PBS, IL-24, low-dose DDP, high-dose DDP, IL-24+low dose DDP and IL-24+high-dose DDP. At the end of the trial, from the conditions of weights and tumor growth to nude mice, PCR technique was tested to the expression of m RNA of IL-24 in transplantation tumor. In addition, there was a detection that whether lymphoid tissue was transplanted tumor lymphatic metastasis by CK keratin. Finally, the expression of m TOR and P70S6 K were tested by immunofluorescence technique as well as Western blot imprinting method to quantitative analysis of its expression. Results IL-24 was successfully transfected and expressed in nude mice xenograft models, the CK keratin for each group was successfully tested in carcinoma infiltrating lymphoid tissues, after intervention, the numbers of metastasis lymph node in different groups were different, that is, the combined group was obvious lower than others(P<0.05), moreover, there was no statistically significant difference(P>0.05) on IL-24+low-dose cisplatin group and IL-24+high-dose cisplatin. m TOR/P70S6 K was successfully tested in all of groups by immunofluorescence technique, and quantified by Western blot analysis, compared with other groups, the combined group was significantly decreased(P<0.05), the difference was statistically significant, and the expression of each two proteins had significant positive correlation(r=0.968, P<0.001). Conclusions The therapeutic effect on IL-24+low-dose DDP group is same with IL-24+high-dose DDP, moreover, IL-24+low-does DDP can significantly inhibit tumor growth and lymph number of metastasis. It is concluded that IL-24 combined with DDP in body inhibit the activity of m TOR/P70S6 K signaling pathway to promote cell apoptosis signaling, from which human cervical cancer cells would be inhibited in nude mice growth and lymph node metastasis.更多还原