热休克蛋白HSPA1A促进肿瘤细胞增殖的研究

Heat Shock Protein A1A Promotes Proliferation of Cancer Cell

摘要:

本研究主要探索热休克蛋白A1A（HSPA1A）对HeLa、HT29、PC3这3种肿瘤细胞增殖的影响及其潜在机制。构建HSPA1A重组载体，转染肿瘤细胞，通过观察转染后细胞形态、MTT测定结果和细胞周期，探索HSPA1A对肿瘤细胞增殖的影响，通过免疫荧光的方法确定HSPA1A对癌细胞增殖发挥作用的细胞定位。过表达HSPA1A后的肿瘤细胞，细胞增殖明显增强，细胞免疫荧光检查确定HSPA1A主要定位于细胞质中，细胞周期实验表明转染HSPA1A主要加快细胞生长周期的G1期，通过Western Blot确定了过表达HSPA1A使得G1期相关的cyclin D1、CDK4、CDK6、cyclin E1和CDK2上调，推测HSPA1A可能作为伴侣蛋白质维持cyclin D1-CDK4/CDK6和（或）cyclin E1-CDK2相关蛋白质的稳定性加快细胞周期的G1期。HSPA1A能够促进肿瘤细胞增殖，且在细胞质中发挥增殖作用。本研究为基于HSPA1A的抗肿瘤药物研发提供了理论依据。

Abstract:

This study was to explore the effects of heat shock protein A1A(HSPA1A) on the proliferation of HeLa, HT29 and PC3 tumor cells and their underlying mechanisms. HSPA1A recombinant vector was constructed to transfect tumor cells. The cell morphology, MTT and cell cycle after transfection were observed to explore the effect of HSPA1A on tumor cell proliferation. The cells that HSPA1A played a role in cancer cell proliferation were determined by immunofluorescence positioning. After overexpression of HSPA1A, cell proliferation of tumor cells was significantly enhanced. Cellular immunofluorescence confirmed that HSPA1A was mainly localized in the cytoplasm. Cell cycle assay showed that transfection of HSPA1A mainly accelerated the G1 phase of cell growth cycle, which was determined by Western Blot. Overexpression of HSPA1A up-regulated G1-related cyclin D1, CDK4, CDK6, cyclin E1, and CDK2. It was speculated that HSPA1A might serve as a chaperone protein to maintain the stability of cyclin D1-CDK4/CDK6 and cyclin E1-CDK2-related proteins and accelerate the G1 phase of cell cycle. HSPA1A can promote tumor cell proliferation and play a role in the cytoplasm proliferation. This study provides a theoretical basis for the development of anti-tumor drugs based on HSPA1A.