摘要
目的 研究重组RA5 38(Ad RA5 38)、反义c myc腺病毒 (Ad ASc myc)在bcl 2高表达细胞系中的作用及其分子机制。方法 采用细胞形态学观察、MTT、RT PCR和Northernblot等方法 ,研究RA5 38、反义c myc重组腺病毒在bcl 2高表达的人宫颈癌细胞系HeLa bcl2和SiHa bcl2以及在其亲本细胞中的生物学作用及其分子机制。结果 以Lipofectin转染bcl 2真核表达质粒建立的bcl 2基因高表达细胞系HeLa bcl2和SiHa bcl2 ,均能高效、稳定地表达bcl 2。Ad RA5 38和Ad ASc myc对HeLa和SiHa均有明显的生长抑制及凋亡诱导作用 ,并可以抑制HeLa细胞bcl 2和c myc基因的表达。但Ad RA5 38、Ad ASc myc对高表达bcl 2的HeLa bcl2和SiHa bcl2细胞bcl 2和c myc基因表达虽有一定抑制作用 ,但对其生长抑制及诱导凋亡的作用轻微 ,未能抵消高表达的bcl 2抑制细胞凋亡的作用。结论Ad RA5 38、Ad ASc myc可能是通过抑制bcl 2及c
Objective To study the effects of recombinant adenovirus with RA538 or antisense c myc insert on cervical cancer cell lines with high expression of bcl 2 gene.Methods Cervical cancer cell lines HeLa and SiHa were transduced with full length bcl 2 cDNA by lipofectin and cell clones with stable expression of bcl 2 were selected. They were then transfected with recombinant adenovirus containing RA538 (Ad RA538), antisense c myc (Ad ASc myc) or LacZ (Ad LacZ) gene. Morphologic and molecular changes of the transfected cancer cells were examined by light microscopy, MTT, RT PCR and Northern blot. Results The bcl 2 cDNA was successfully transferred into HeLa and SiHa cells. Two cell lines, called HeLa bcl2 and SiHa bcl2 with high expression of bcl 2 gene were generated. Ad RA538 and Ad ASc myc transfection both could inhibit cell growth and induce apoptosis of HeLa and SiHa cells, and inhibit their expression of c myc and bcl 2 genes. However, although Ad RA538 and Ad ASc myc had some inhibitory effect on bcl 2 and c myc gene expression by HeLa bcl2 and SiHa bcl2 cells, they could only slightly inhibit cell growth and weakly induce apoptosis.Conclusion Ad RA538 and Ad ASc myc can to certain extent inhibit cell growth and induce apoptosis of two cervical cancer cell lines. They have little such effect when these cell lines have over expressed bcl 2 by transduction of exogenous bcl 2 gene.
出处
《中华肿瘤杂志》
CSCD
北大核心
2000年第4期279-282,共4页
Chinese Journal of Oncology
基金
国家 8 6 3高科技发展基金资助项目 !(Z2 0 0 1 0 2 )
