论文部分内容阅读
目的:在前期体内实验(文章已经发表)证实上皮-间质细胞转化核转录因子twist、snail在肺岩宁方抗肿瘤转移具有重要作用的基础上,进一步体外实验观察肺岩宁方对人高转移95-D肺癌细胞侵袭能力及twist、snail及Sip1基因和蛋白表达的影响。方法:应用不同浓度肺岩宁含药血清干预治疗人高转移肺癌细胞95-D的基础上,采用Tran-swell小室法检测细胞侵袭能力,并应用real-time PCR及Western blot方法检测对上皮-间质细胞转化核转录因子twist、snail及Sip1基因和蛋白表达的影响。结果:Transwell上室加入不同浓度肺岩宁方含药血清后,15%、20%及25%的肺岩宁含药血清的穿膜数量明显低于对照组(P<0.01);Real-time PCR结果表明:与对照组相比较,不同浓度肺岩宁含药血清对Twist均有下调作用(P<0.01),5%、10%、15%浓度的含药血清及DDP对Snail有下调作用(P<0.01);10%、15%肺岩宁方含药血清及DDP对Sip1有下调作用(P<0.05或P<0.01)。Western blot结果表明:5%、10%肺岩宁方含药血清具有显著下调Twist蛋白表达的作用(P<0.01),5%、15%浓度的肺岩宁方含药血清对Snail有下调作用(P<0.01),15%肺岩宁方含药血清对Sip1蛋白有下调作用(P<0.01)。结论:肺岩宁方通过抑制Twist、Snail及SIP1的表达,从而抑制肿瘤异质黏附能力和运动能力来参与肺癌的侵袭和转移。
OBJECTIVE: In the previous in vivo experiments (articles published) confirmed that epithelial-mesenchymal transition of nuclear transcription factor twist, snail in the anti-tumor lung-yan Fang metastasis on the basis of an important role in further experiments in vitro Feiyanning on human high Metastasis of 95-D lung cancer cells and the expression of twist, snail and Sip1 genes and proteins. Methods: The cell invasion was detected by Tran-swell chamber method based on the intervention of different concentrations of Feiyanning serum in human high metastatic lung cancer cell line 95-D. Real-time PCR and Western blotting were used to detect the expression of epithelial- Effects of interstitial cell transforming transcription factor twist, snail and Sip1 gene and protein expression. RESULTS: After the different concentrations of Feiyanningfang serum were added into the upper chamber of Transwell, the numbers of penetrating membrane of the drug-containing serums of Feiyanning were significantly lower than those of the control group (P <0.01). Real-time PCR results showed that compared with the control group, different concentrations of Feiyanning serum had a downward effect on Twist (P <0.01), 5%, 10%, 15% concentration of drug-containing serum and DDP down-regulation of Snail (P <0.01). Sip1 was down-regulated by 10% and 15%, respectively (P <0.05 or P <0.01). The Western blot results showed that 5% and 10% of Feiyanning decoction serum could significantly down-regulate the expression of Twist protein (P <0.01), while 5% and 15% of Feiyanning decoction serum could down-regulate Snail (P <0.01). 15% Feiyanningfang serum reduced Sip1 protein (P <0.01). Conclusion: Feiyanning Decoction can participate in the invasion and metastasis of lung cancer by inhibiting the expression of Twist, Snail and SIP1, thereby inhibiting the heterogeneous adhesion and movement ability of tumor.