LASS2表达对人膀胱癌细胞EJ凋亡和生物学行为的影响及机制

The main research objectives are to learn the function and mechanism of Homo sapiens LAG1 longevity assurance 2 on human bladder cancer EJ cell line. In this study, expression vectors, cell transfection and other experimental methods were used to establish a high-expression cell line expressing LASS2 gene. By means of siRNA methods, we will screen out a cell subline without expressing LASS2 gene. Apoptosis rates of the three cell lines will be detected by using the method of flow cytometry. Effect of LASS2 expression on the abilities of cell proliferation, adhesion, invasion and metastasis will be explicit. Cytoskeleton structural changes will be observed by cytoskeleton staining, oil and scanning electron microscopy. Expression of Ezrin will be detected by Real-time quantitative PCR and Western Blot. Differences of telomerase activity will be detected by TRAP-ELISA method. This study was designed to study the anticancer effect of LASS2 and to evaluate its latent energy of LASS2 in prevention and clinical application of human bladder cancer.

本项目主要研究人源性长寿保障基因Ⅱ型LASS2在膀胱癌细胞EJ中的功能和作用机理：通过表达载体构建，细胞转染等实验方法建立LASS2稳定高表达的膀胱癌EJ细胞株；通过siRNA沉默等实验手段筛选出LASS2稳定不表达的膀胱癌EJ细胞株。以筛选出的两株细胞系和母代的膀胱癌EJ细胞为研究对象，运用流式细胞术检测三株细胞的凋亡率和细胞周期；通过体外和体内实验明确LASS2表达对膀胱癌EJ细胞增殖能力、黏附能力、侵袭和转移能力及其相关因子表达的影响；通过细胞骨架染色，油镜和扫描电镜观察细胞骨架结构变化，实时定量PCR、Western Blot检测细胞骨架连接分子Ezrin的表达差异，TRAP-ELISA法检测各组细胞端粒酶活性的异同，研究LASS2因子对膀胱癌细胞的作用机理，以期更好地发掘LASS2的抗癌作用，评估LASS2应用于膀胱癌预防和临床治疗的潜质。

本项目主要研究人源性长寿保障基因Ⅱ型LASS2 在膀胱癌细胞EJ 中的功能和作用机理：通过表达载体构建，细胞转染等实验方法建立LASS2 稳定高表达的膀胱癌EJ 细胞株；通过siRNA 沉默等实验手段筛选出LASS2 稳定不表达的膀胱癌EJ 细胞株。以筛选出的两株细胞系和母代的膀胱癌EJ 细胞为研究对象，运用流式细胞术检测三株细胞的凋亡率和细胞周期；通过体外和体内实验明确LASS2 表达对膀胱癌EJ 细胞增殖能力、黏附能力、侵袭和转移能力及其相关因子表达的影响，并探讨相关机制。通过上述实验证明了LASS2基因可能参与了膀胱癌EJ细胞增殖、迁移和细胞周期变化的过程，为进一步的实验打下基础，并对其用于膀胱癌诊断和治疗的潜在前景做出了评估。