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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 chemosensitizing effect and its molecular mechanism of ING4 (costimulatory factor 4) and IL-24 (human interleukin 24) double-gene adenovirus vector (Ad-ING4-IL-24) on lung adenocarcinoma. The effects of Ad-ING4-IL-24 on A549 lung cancer cells and cisplatin (DDP) chemotherapeutic drugs were detected. The transcription and expression of ING4 and IL-24 genes in A549 lung cancer cells were detected by RT-PCR and Western blotting. MTT assay The growth inhibition and apoptosis effects of Ad-ING4-IL-24 combined with DDP (combination group) on A549 lung cancer cells were detected by FCM and Hoechst staining. The A549 cell line was used to establish the human lung adenocarcinoma model in nude mice. The tumor volume was measured by local injection and the tumor volume was measured dynamically. The tumor inhibition rate was calculated. The expressions of ING4, IL-24, bax, bcl-2, VEGF and other genes expression. The results showed that Ad-ING4-IL-24 could be successfully transcribed and expressed in A549 lung cancer cells. The combination of Ad-ING4-IL-24 and A549 could significantly inhibit the growth of A549 cells and induce the apoptosis of A549 cells. In vivo combination group also can significantly inhibit tumor growth, tumor weight inhibition rate was 52.81%, immunohistochemistry results showed that the combination group can up-regulate bax gene expression, while down-regulating bcl-2, VEGF and other gene expression. These results suggest that Ad-ING4-IL-24 has chemosensitizing effect, which may be related to promoting tumor cell apoptosis and inhibiting angiogenesis.