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目的:探讨六味地黄丸联合基因治疗对小鼠移植性肝癌缝隙连接蛋白(connexin,Cx)表达的影响,初步阐明六味地黄丸对肝癌HSV-tk/GCV自杀基因治疗的增效作用机制是否与缝隙连接相关。方法:昆明种小鼠90只,随机分为模型对照组、六味地黄丸治疗组(ig六味地黄丸10 g·kg-1·d-1)、自杀基因治疗组(ip丙氧鸟苷100 mg·kg-1·d-1)、联合治疗组(ig六味地黄丸10 g·kg-1.d-1+ip丙氧鸟苷100 mg·kg-1.d-1);选用H22与H22/tk细胞株皮下接种建立小鼠肝癌模型,取荷瘤小鼠肿瘤组织进行常规病理制片,采用SABC免疫组化法检测各组肿瘤组织Cx26,Cx32,Cx43蛋白表达。结果:阳性表达定位于细胞浆和细胞膜,Cx43在癌旁组织,特别是有纤维组织浸润部位表达呈强阳性,而在癌结节中肿瘤细胞表达相对较弱;Cx26,Cx32在肿瘤细胞特别是脂肪组织浸润附近的癌组织中呈现强阳性表达。与肿瘤模型组相比,联合治疗组则能增强Cx26,Cx43,Cx32蛋白表达,差异有统计学意义(P<0.05);与单独六味地黄丸和自杀基因治疗组相比,联合治疗组能明显提高Cx32蛋白表达,差异有统计学意义(P<0.05,P<0.01)。结论:六味地黄丸联合自杀基因治疗能增强Cx26,Cx43,Cx32蛋白表达,尤其是促进Cx32的表达,提示六味地黄丸增强自杀基因旁观者效应的机制与缝隙连接相关。
Objective: To investigate the effect of Liuweidihuangwan combined gene therapy on the expression of connexin (Cx) in transplanted hepatocellular carcinoma in mice and to clarify whether the synergistic mechanism of Liuweidihuang Pill on HSV-tk / GCV suicide gene therapy is related to the gap Connection related. Methods: Ninety Kunming mice were randomly divided into four groups: model control group, Liuweidihuangwan treatment group (ig, Liuweidihuangwan 10 g · kg -1 · d -1), suicide gene therapy group (ip ganosine 100 mg Kg-1 · d-1), combined treatment group (ig, Liuweidihuangwan 10 g · kg -1 · d -1 + ip ganoside 100 mg · kg -1 · d -1), H22 and H22 / tk cells were inoculated subcutaneously to establish a mouse model of hepatocellular carcinoma. The tumor tissues of the tumor-bearing mice were prepared for routine pathology. The expression of Cx26, Cx32 and Cx43 in each tumor tissue was detected by SABC immunohistochemistry. Results: The positive expression was located in the cytoplasm and cell membrane. The expression of Cx43 was strongly positive in the paracancerous tissues, especially in the infiltrated fibrous tissue, while the expression of Cx26 and Cx32 was relatively weak in the tumor nodules, especially in the tumor cells Adipose tissue infiltration in the vicinity of the strong positive expression of cancer tissue. Compared with the tumor model group, the combination therapy group can enhance the Cx26, Cx43, Cx32 protein expression, the difference was statistically significant (P <0.05); compared with the Liuweidihuangwan and suicide gene therapy group, the combination therapy group can be significantly increased Cx32 protein expression, the difference was statistically significant (P <0.05, P <0.01). Conclusion: Liuweidihuangwan combined with suicide gene therapy can enhance the expression of Cx26, Cx43 and Cx32 protein, especially promote the expression of Cx32, suggesting that the mechanism of Liuweidihuangwan enhancing the bystander effect of suicide gene is related to gap junction.