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目的探讨RNA干扰(RNA interference,RNAi)生存素(Survivin)基因联合X射线照射对裸鼠移植人肝癌细胞SMMC-7721的抑瘤效应。方法肿瘤局部注射脂质体包裹的靶向Survivin基因的RNA干扰质粒后,接受5Gy X射线照射,观察各组裸鼠治疗后不同时间肿瘤体积和平均存活时间,以免疫组化染色法检测肿瘤组织Survivin、增殖细胞核抗原(Proliferation Cell Nuclear Antigen,PCNA)表达和肿瘤间质微血管密度,以TUNEL法检测肝癌细胞凋亡。结果 治疗后第3～21d,RNA干扰联合放疗组肿瘤体积明显低于对照组、RNA干扰组和放疗组。RNA干扰联合放疗组裸鼠平均存活时间明显长于对照组、RNA干扰组和放疗组。治疗结束后第1天,RNA干扰联合放疗组裸鼠肿瘤增殖活性和微血管密度明显低于RNA干扰组和放疗组,且凋亡细胞百分数明显升高。结论 RNA干扰Survivin基因联合放疗可有效地抑制裸鼠移植人肝癌细胞增殖和血管生成,促进细胞凋亡,其抑瘤效应明显优于放疗和RNA干扰治疗。
Objective To investigate the antitumor effect of RNA interference (RNAi) survivin gene combined with X-ray irradiation on human hepatocellular carcinoma cell line SMMC-7721. Methods The tumor-bearing mice were injected with liposome-encapsulated RNA interference plasmids targeting Survivin gene and then irradiated with 5Gy X-ray. The tumor volume and mean survival time were observed at different time points after treatment in nude mice. The tumor tissues were detected by immunohistochemical staining Survivin, Proliferation Cell Nuclear Antigen (PCNA) expression and tumor interstitial microvessel density were detected by TUNEL assay. Results The tumor volume of RNA interference combined with radiotherapy group was significantly lower than that of the control group, RNA interference group and radiotherapy group from the 3rd to 21st days after treatment. The mean survival time of nude mice in RNA interference combined with radiotherapy group was significantly longer than that in control group, RNA interference group and radiotherapy group. On the first day after treatment, the tumor proliferation activity and microvessel density in nude mice treated with RNA interference and radiotherapy were significantly lower than those in RNA interference and radiotherapy groups, and the percentage of apoptotic cells was significantly increased. Conclusion Survivin RNA interference combined with radiotherapy can effectively inhibit the proliferation and angiogenesis of human hepatocellular carcinoma cells in nude mice and promote apoptosis. The antitumor effect is better than radiotherapy and RNA interference therapy.