Ad-ING4-IL-24双基因共表达对人肺腺癌化疗的增敏效应

Effect of adenovirus-mediated ING4 and IL-24 co-expression on chemosensitivity to human lung adenocarcinoma in vitro and in vivo

研究ING4(肿瘤生长抑制因子4)和IL-24(人白细胞介素24)双基因共表达腺病毒载体(Ad-ING4-IL-24)对肺腺癌的化疗增敏效应及分子机制。将Ad-ING4-IL-24感染A549肺癌细胞及联合顺铂(DDP)化疗药物作用,RT-PCR和Western blotting检测ING4和IL-24基因在A549肺癌细胞中的转录和表达,MTT法、流式细胞仪(FCM)和Hoechst染色法检测Ad-ING4-IL-24联合DDP(联合组)对A549肺癌细胞的生长抑制和凋亡效应。采用A549细胞株建立人肺腺癌裸鼠模型,然后瘤体局部注射干预用药,动态测量肿瘤体积,并计算瘤重抑瘤率,免疫组化检测ING4、IL-24、bax、bcl-2、VEGF等基因的表达。结果表明,Ad-ING4-IL-24感染A549肺癌细胞后可获得成功转录和表达,体外联合组能明显抑制A549肺癌细胞生长和诱导细胞凋亡,呈现出典型细胞凋亡形态学变化。体内联合组同样能显著抑制肿瘤生长,瘤重抑瘤率达52.81%,免疫组化结果显示联合组能上调bax基因表达,同时下调bcl-2、VEGF等基因的表达。以上结果表明Ad-ING4-IL-24具有化疗增敏的作用,该作用机制可能与促进肿瘤细胞凋亡和抑制血管形成有关。

To study the chemosensitivity and the mechanisms of recombinant adenovirus vector expressing ING4 and IL-24（Ad-ING4-IL-24） on lung adenocarcinoma in vitro and in vivo,the expression of ING4 and IL-24 in A549 cells was detected by RT-PCR and Western blotting.The growth inhibition,apoptosis rate and apoptosis body were measured by MTT,flow cytometry and Hoechst staining respectively.For in vivo study,we first established the A549 tumor model by grafting A549 cells in athymic nude mice;and then injected Ad-ING4-IL-24 into the tumors.Two weeks after injection,we killed the mice,removed the tumors,weighted and calculated the ratios of tumor-suppression.We also detected the expressions of ING4,IL-24,bax,bcl-2,VEGF with immunohistochemistry.The results indicated that ING4 and IL-24 were proved successfully transcription and expression in A549 cells.More interestingly,the joint group inhibited the growth of A549 cells and induced apoptosis.The in vivo data showed that the joint group suppressed the tumor growth conspicuously through up-regulating the expression of bax,and down-regulating the expression of bcl-2,VEGF.The study proved that Ad-ING4-IL-24 significantly enhanced the chemosensitivity to anticancer drug DDP in lung adenocarcinoma,which may related with cell apoptosis and antiangiogenesis.