论文部分内容阅读
目的:通过检测参芪扶正注射液对肝癌MHCC97L细胞侵袭力及其VEGF基因表达的影响,明确其降低肝癌瘤株侵袭力的作用,并通过RNA干扰技术加以验证。方法:在MHCC97L体外培养系中加入参芪扶正注射液(0.5 g/L)作用24 h后,Transwell检测参芪扶正注射液对MHCC97L细胞侵袭性的影响;同时使用Real-time PCR和Western-blot分别检测药物组、阴性对照组和空白对照组中VEGF基因的转录与表达情况,确定参芪扶正注射液对VEGF基因转录与表达的影响;构建VEGF干扰慢病毒载体并实现其在人肝癌细胞MHCC97L中表达,并通过RT-PCR、Western-Blot进行验证,确定其干扰VEGF基因表达的有效性;Transw ell检测参芪扶正注射液对VEGF“沉默”瘤株侵袭力的影响并与正常瘤株进行比较。结果:参芪扶正注射液能够降低MHCC97L细胞中VEGF基因的转录与表达(P<0.05),并能够降低肿瘤侵袭力(P<0.05);VEGF基因“沉默”瘤株构建成功;VEGF基因“沉默”后,参芪扶正注射液对MHCC97L瘤株侵袭力的影响不显著(P>0.05)。结论:参芪扶正注射液可以通过抑制MHCC97L细胞VEGF基因的表达以降低其侵袭力。
OBJECTIVE: To determine the effect of Shenqi Fuzheng injection on invasiveness of human hepatocellular carcinoma (MHCC97L) cells and VEGF gene expression, and to clarify the effect of Shenqi Fuzheng injection on reducing invasiveness of hepatocellular carcinoma. Methods: The effects of Shenqi Fuzheng injection on the invasiveness of MHCC97L cells were detected by Transwell assay after adding Shenqi Fuzheng Injection (0.5 g / L) for 24 h in MHCC97L in vitro culture system. Real-time PCR and Western-blot The transcription and expression of VEGF gene in drug group, negative control group and blank control group were detected respectively to determine the effect of Shenqi Fuzheng injection on the transcription and expression of VEGF gene. VEGF interference lentiviral vector was constructed and its expression in human hepatoma cell line MHCC97L Transient assay was used to detect the effect of Shenqi Fuzheng injection on the invasiveness of VEGF “Silencing ” tumor cells and compared with the normal tumor Strains were compared. Results: Shenqi Fuzheng injection could reduce the transcription and expression of VEGF gene in MHCC97L cells (P <0.05), and could reduce the invasiveness of tumor (P <0.05). VEGF gene silencing was successfully constructed. VEGF gene “Silence ”, Shenqi Fuzheng injection had no significant effect on invasiveness of MHCC97L (P> 0.05). Conclusion: Shenqi Fuzheng injection can reduce the invasion of MHCC97L cells by inhibiting the expression of VEGF gene.