摘要
目的 探讨BAK基因过表达对膀胱癌细胞的诱导凋亡效应及机制。方法 脂质体介导BAK基因转染膀胱癌EJ细胞 1～ 7d后 ,逆转录聚合酶链反应检测BAK基因表达 ,细胞计数法检测癌细胞生长活性 ,DNALadder法、吖啶橙 溴化乙锭荧光染色法及原位末端转移酶标记技术检测癌细胞凋亡 ;免疫组织化学法检测癌细胞Caspase 3、Bcl 2、p5 3表达。 结果 转染 1～ 7d后 ,癌细胞BAK基因表达显著增强 ( P <0 .0 1) ,体外生长抑制 2 0 .6 6 %～ 35 .5 8% ( P <0 .0 1) ;部分癌细胞呈现凋亡形态学变化 ,凋亡率为 18.0 %～ 2 0 .6 % (P <0 .0 1) ;癌细胞Caspase 3表达增强 6 .6倍(P <0 .0 1) ,Bcl 2和 p5 3表达差异无显著性 ( P >0 .0 5 )。结论 BAK基因过表达能显著诱导膀胱癌细胞凋亡 ,其中Caspase 3激活是其作用机制之一。
Objective To explore the apoptosis induction of bladder cancer cells by BAK gene transfer and its mechanism.Methods After being transferred by BAK gene with liposome for 1 to 7 days,the BAK gene expression level in cancer cells was detected by reverse transcription polymerase chain reaction (RT-PCR).The growth activities of cancer cells were detected by cell count method.Apoptosis was assayed by DNA ladder,acridine orange-ethidium bromide fluorescent staining,and in situ terminally labeled transferase technique (TUNEL).The cellular expression of Caspase-3,Bcl-2 and p53 were observed by immunohistochemistry method.Results After being transferred for 1 to 7 days,the BAK expression wasincreased significantly (P<0.01).The growth of bladder cancer cells was decreased by 20.66 % to 35.58 % (P<0.01).Partial cancer cells presented the characteristic morphological changes of apoptosis with the apoptosis rates being 18.0 % to 20.6 % (P<0.01).The cellular expression of Caspase-3 was elevated by 6.6 times.There were no significant changes in the cellular expression of Bcl-2 and p53 (P>0.05).Conclusion BAK gene over-expression could induce apoptosis of bladder cancer cells.The activation of Caspase-3 was one of its mechanisms.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2003年第1期63-64,共2页
Chinese Journal of Experimental Surgery
