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Suppression effect of recombinant adenovirus vector containing hIL-24 on Hep-2 laryngeal carcinoma cells

机译:含hIL-24重组腺病毒载体对Hep-2喉癌细胞的抑制作用

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摘要

The melanoma differentiation-associated gene-7 [MDA-7; renamed interleukin (IL)-24] was isolated from human melanoma cells induced to terminally differentiate by treatment with interferon and mezerein. MDA-7/IL-24 functions as a multimodality anticancer agent, possessing proapoptotic, antiangiogenic and immunostimulatory properties. All these attributes make MDA-7/IL-24 an ideal candidate for cancer gene therapy. In the present study, the human MDA-7/IL-24 gene was transfected into the human laryngeal cancer Hep-2 cell line and human umbilical vein endothelial cells (HUVECs) with a replication-incompetent adenovirus vector. Reverse transcription polymerase chain reaction and western blot analysis confirmed that the Ad-hIL-24 was expressed in the two cells. The expression of the antiapoptotic gene, Bcl-2, was significantly decreased and the IL-24 receptor was markedly expressed in Hep-2 cells following infection with Ad-hIL-24, but not in HUVECs. In addition, the expression of the proapoptotic gene, Bax, was induced and the expression of caspase-3 was increased in the Hep-2 cells and HUVECs. Methyl thiazolyl tetrazolium assay indicated that Ad-hIL-24 may induce growth suppression in Hep-2 cells but not in HUVECs. In conclusion, Ad-hIL-24 selectively inhibits proliferation and induces apoptosis in Hep-2 cells. No visible damage was found in HUVECs. Therefore, the results of the current study indicated that Ad-hIL-24 may have a potent suppressive effect on human laryngeal carcinoma cell lines, but is safe for healthy cells.
机译:黑色素瘤分化相关基因7 [MDA-7;从人黑素瘤细胞中分离出重命名的白介素(IL)-24],该黑素瘤细胞经干扰素和美黑素的处理最终分化。 MDA-7 / IL-24用作多模态抗癌药,具有促凋亡,抗血管生成和免疫刺激特性。所有这些属性使MDA-7 / IL-24成为癌症基因治疗的理想候选者。在本研究中,使用无复制能力的腺病毒载体将人MDA-7 / IL-24基因转染到人喉癌Hep-2细胞系和人脐静脉内皮细胞(HUVEC)中。逆转录聚合酶链反应和蛋白质印迹分析证实,Ad-hIL-24在两个细胞中表达。 Ad-hIL-24感染后,Hep-2细胞中抗凋亡基因Bcl-2的表达显着降低,IL-24受体显着表达,但HUVEC中未表达。另外,在Hep-2细胞和HUVEC中诱导了凋亡基因Bax的表达,并且胱天蛋白酶3的表达增加。甲基噻唑基四唑鎓测定表明,Ad-hIL-24可能在Hep-2细胞中诱导生长抑制,但在HUVECs中不诱导生长抑制。总之,Ad-hIL-24在Hep-2细胞中选择性抑制增殖并诱导凋亡。在HUVEC中未发现可见的损坏。因此,本研究的结果表明,Ad-hIL-24可能对人喉癌细胞系具有有效的抑制作用,但对健康细胞是安全的。

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