摘要
目的探讨连接蛋白基因Cx43对胶质瘤细胞增殖的抑制及其可能的机理。方法将含Cx43cDNA的质粒以脂质体介导转染Cx43表达缺失的人和鼠的恶性胶质瘤细胞,通过Northem杂交、原位杂交及免疫组化染色检测Cx43mRNA及蛋白表达;MTT法测定细胞增殖率;核仁组成区嗜银蛋白染色检测细胞增殖活性;TUNEL法检测细胞凋亡;划痕标记荧光染料示踪技术检测细胞间隙连接通讯(GJIC);Western杂交及免疫组化染色检测bFGF、PDGF、EGFR、IGF-Ⅰ和IGFBP3的表达。结果转染Cx43基因的胶质瘤细胞增殖下降,GJIC恢复,同时伴有bFGF、PDGF、IGF-Ⅰ和IGFBP3表达下降,而EGFR表达和细胞凋亡则无改变。结论Cx43基因可能通过恢复GJIC功能及抑制某些重要生长因子的自分泌,实现对胶质瘤细胞增殖的抑制。
Objective To study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene. Methods C6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-1 and IGFBP3 expression. Results Cx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change. Conclusion The mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2003年第1期4-8,共5页
Chinese Journal of Oncology
基金
国家自然科学基金资助项目(39870815)
