Ad-PTEN对人小细胞肺癌NCI-H446细胞生长的抑制作用  

Inhibition Effect of Adenovirus-mediated PTEN Gene on NCI-H446 Cell Growth

作　　者：李东蓉[1] 杨吉成[2] 盛伟华[2] 谢宇锋[2] 朱晔涵[1] 

机构地区：[1]苏州大学附属第一医院呼吸内科,江苏苏州215006 [2]苏州大学医学部基础医学与生物科学学院细胞生物学系,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2011年第4期522-526,共5页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金资助项目(81001016);江苏省卫生厅卫生科技项目资助(H200914)

摘　　要：目的研究重组腺病毒第10号染色体同源丢失性磷酸酶张力蛋白基因(PTEN)对小细胞肺癌的抑癌增效和化疗增敏效应及其分子机制。方法采用重组腺病毒载体PTEN(简称Ad-PTEN)感染QBI-293A细胞,并用该细胞进行病毒扩增和效价测定。将Ad-PTEN感染NCI-H446小细胞肺癌细胞。分为5组:PBS组、空载体Ad组、Ad-PTEN组、顺铂(DDP)组和Ad-PTEN+DDP组,Western blot鉴定PTEN基因表达,MTT法和流式细胞仪(FCM)检测Ad-PTEN对NCI-H446小细胞肺癌细胞的生长抑制和诱导凋亡的增效作用,用RT-PCR检测细胞凋亡相关基因p53、bax、caspase-3、bcl-2、和survivin的转录。结果 Ad-PTEN可在QBI-293A细胞中增殖。Ad-PTEN感染NCI-H446细胞后,Western blot检测到了PTEN基因的表达。Ad-PTEN组、DDP组、Ad-PTEN+DDP组体外能明显抑制人NCI-H446小细胞肺癌细胞的生长和诱导凋亡,且Ad-PTEN+DDP组效应较Ad-PTEN组、DDP组更为显著(均P<0.05)。Ad-PTEN组、DDP组、Ad-PTEN+DDP组的NCI-H446细胞中的p53、bax及caspase-3基因表达均上调,而Bcl-2和Survivin基因表达均下调,这些凋亡相关基因的上调或下调的表达趋势以Ad-PTEN+DDP组最显著(均P<0.05)。结论 Ad-PTEN体外可抑制NCI-H446小细胞肺癌细胞生长,Ad-PTEN对DDP的细胞毒作用具有增敏效应,其机制可能与上调p53、bax及caspase-3基因表达和下调bcl-2和survivin基因表达有关。Objective To explore effect of inhibitory synergy and chemosensitivity of recombinant adenovirus phosphatase and tensin homologue deleted chromatosome10（PTEN） on small lung cancer and its possible mechanisms.Methods Recombinant adenovirus Ad-PTEN was constructed and transfected into the human embryonic kidney 293（QBI-293A） cells.The adenovirus completed the amplification in QBI-293A cells and the titration was detected.The human pulmonary carcinoma cell line NCI-H446 was infected by Ad-PTEN and combined with cisplatin（DDP） in vitro,and were randomly divided into five groups：the PBS negative control group（PBS group）,the idling adenovirus group（Ad group）,the adenovirus-mediated PTEN group（Ad-PTEN group）,the DDP positive control group（DDP group） and the group of Ad-PTEN combined with DDP（referred to as joint group）.The expression of PTEN in NCI-H446 small cell lung cancer cells was detected by Western blot.Effect of enhanced growth-suppressing and apoptosis-inducing by Ad-PTEN on NCI-H446 small cell lung cancer cells was analyzed by MTT assay and Flow cytometry.Expression of apoptosis-related genes（p53、bax、caspase-3、bcl-2、survivin） was detected by reverse transcription-PCR（RT-PCR）.Results After the amplification of Ad-PTEN in QBI-293A cells,the titration was 108pfu/mL.The expressions of PTEN in NCI-H446 cells were detected by Western blot.Adenovirus-mediated PTEN significantly inhibited NCI-H446 cell growth,induced cell apoptosis in vitro.The p53,bax,and caspase-3 expression of NCI-H446 cells significantly increased in the Ad-PTEN group,DDP group,Ad-PTEN＋DDP group,while Bcl-2 and Survivin gene expression significantly reduced;and in the Ad-PTEN＋DDP group it was most significant（P0.05）.Conclusion Ad-PTEN can inhibit small cell lung cancer NCI-H446 cells in vitro,Ad-PTEN to DDP cytotoxicity with a sensitizing effect.Its mechanism may be related to increaseing the expression level of p53、 bax and caspase-3 and reducing the expression level of bcl-2 and survivi

关 键 词：第10号染色体同源丢失性磷酸酶张力蛋白基因 小细胞肺癌 抑癌效应 化疗增敏 

分 类 号：R734.2[医药卫生—肿瘤]