槐耳清膏对人高转移大细胞肺癌细胞L9981血管生成相关基因表达的影响

Effects of polysaccharid on expression of angiogenic-related genes in human high-metastatic large cell lung cancer cell line L9981

背景与目的肺癌是全世界对人类健康与生命危害最大的恶性肿瘤之一。目前从植物中开发抗肿瘤药物是国内外抗肿瘤新药开发的一个热点。本研究的目的是观察槐耳清膏对人高转移大细胞肺癌细胞株L9981血管生成相关基因mRNA转录表达的影响及意义,并探讨其可能的分子机制。方法体外培养人高转移大细胞肺癌细胞L9981,应用台盼蓝拒染法做细胞毒性实验,实验分为:空白对照组,槐耳清膏组,顺铂化疗组,联合用药组。应用RTPCR检测四组细胞株用药前后βcatenin、Ecadherin、TIMP1、CD44V6、MMP2、endostatin、VEGFmRNA的表达。应用Boyden小室法检测四组细胞株用药前后体外侵袭力改变。结果①槐耳清膏对L9981细胞的生长有明显的抑制作用,且抑制率随药物浓度增加而上升。槐耳清膏组(1g/L)与顺铂化疗组(顺铂3mg/L)及联合用药组(槐耳清膏0.05g/L+顺铂1.5mg/L)比较抑制率无明显差异(P>0.05)。②经槐耳清膏处理后,L9981细胞的βcatenin、Ecadherin、TIMP1、endostatin、MMP2的mRNA表达水平上调,而VEGF、CD44V6表达水平下调,其中TIMP1、endostatin表达水平明显上调,CD44V6表达水平明显下调。③槐耳清膏组、顺铂化疗组、联合用药组细胞株的体外侵袭力均明显低于空白对照组(P<0.05)。结论①槐耳清膏在体外对人高转移大细胞肺癌细胞L9981具有生长抑制作用,该抑制作用呈剂量依赖性。②槐耳清膏对L9981的生长抑制作用可能与其调控血管生成相关基因mRNA的表达有关。③槐耳清膏联合顺铂有一定的协同抗肿瘤作用。

Background and objective Lung cancer is one of the most malignant cancers which is hazarding the people＇s health and life in the world. At present, it is a highlight to exploit antitumor drug from plant at home and abroad. The aim of this study is to observe the effects of polysaccharid （PS-T） on expression of angiogenic-related gene mRNA in human high-metastatic large cell lung cancer cell line L9981, and to explore its possible molecular mechanism. Methods L9981 in vitro was cultured, and the growth data were obtained by trypan blue staining. The mRNA transcript expression of β-catenin, E-cadherin, TIMP-1, CD44V6, MMP-2, endostatin, VEGF was detected in L9981 by RT-PCR before and after treating with PS-T. The ability of invasion of L9981 was determined by Boyden chamber method. Results OPS-T had remarkably inhibitive effects on the growth of L9981 in vitro. The inhibitive rate of PS-T on L9981 was concentration-depend- ent. No significant difference of inhibitive rate was found among the PS-T （1 g/L）, cisplatin （3 mg/L） and PS-T （0.05 g/L） ＋ cisplatin （1.5 mg/L）（P〉0. 05）. ①The mRNA expression level of β-catenin, E-cadherin, TIMP-1, endostatin and MMP-2 was upregulated, while that of VEGF and CD44V6 was downregulated. Out of them the mRNA expression level of TIMP-1 and endostatin was remarkably upregulated, the expression level of CD44V6 was significanyly downregulated. ③The in vitro invasive abilities of L9981 was significantly decreased in the PS-T, DDP and PS-T-HDDP groups compared with that in blank control group. Conclusion ①PT-S could inhibit the growth of human high metastatic large cell lung cancer cell line L9981 in vitro, the effect is dose-dependent.②PS-T can down- or up-regulate the mRNA transcript expression of some angiogenic-related gene mRNA. ③PS-T has remarkably coordinating effects with cisplatin in the L9981 lung cancer cell line.