病毒介导IL-24基因对口腔鳞癌耐药细胞株凋亡的影响  

Virus Vector-mediated IL-24 Induces Apoptosis in Drug-resistant Oral Squamous Cell Carcinoma KBV Cells

作　　者：杨德钊[1] 王丹丹[2] 孟祥[1] 杨帅[1] 孙宏晨[2] 李吉辰[1] 

机构地区：[1]哈尔滨医科大学附属口腔医院口腔颌面外科,黑龙江哈尔滨150001 [2]吉林大学口腔医院病理科,吉林长春130021

出　　处：《口腔颌面外科杂志》2015年第4期246-252,共7页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金委员会批准资助对外交流与合作项目(30740420551);留学归国科学基金(LC2009C04)

摘　　要：目的:研究杂合病毒介导的IL-24基因对口腔鳞状细胞癌耐药细胞的凋亡诱导作用,探讨其治疗耐药口腔癌的可能性。方法:通过MTT细胞活力测定鉴定口腔鳞状细胞癌细胞株KB和其耐药细胞株KBV。利用杂合病毒Ad LTR2EF1α搭载EGFP和IL-24转染KB细胞,确定最佳转染浓度,荧光倒置显微镜及RT-PCR鉴定EGFP或IL-24基因表达。以人永生化上皮细胞Ha Ca T细胞作为对照,使用Ad LTR2EF1α-IL-24转染KB、KBV及Ha Ca T细胞后,MMT法检测细胞活力,Annexin V/PI双染法检测细胞凋亡,透射电镜观察细胞凋亡形态。结果:KBV细胞对长春新碱的耐受性明显高于KB细胞。Ad LTR2EF1α-IL-24搭载的IL-24基因可以在KB及KBV细胞内过表达。IL-24基因在KB和KBV细胞中的过表达,有明显抑制细胞生长和促凋亡作用,而在Ha Ca T细胞中没有类似的作用。结论 :杂合病毒介导的IL-24对口腔鳞状细胞癌耐药细胞有较强的细胞毒性和诱导凋亡作用,并且对正常组织细胞没有影响。具有治疗耐药口腔癌的潜在价值。Objective： To study the apoptosis-inducing effect of hybrid virus vector-mediated IL-24 gene on oral squamous cell carcinoma（OSCC） cell line and drug-resistant cell line. Methods： In order to identify OSCC cell line KB and the drug resistant counterparts cell lines KBV, cell viability was measured by MTT assay. Fluorescence microscopy and RT-PCR experiments were used to detect the gene expression, Ad LTR2EF1α-EGFP or Ad LTR2EF1α-IL-24 were used to treat the KB cells in order to obtain the optimal drug concentration. Human keratinocyte Ha Cat cells were used as control cells.Ad LTR2EF1α-IL-24 and Ad LTR2EF1α-vec transfected the KB,KBV, and Ha Cat cells in a 200 μM substrate solution, and incubated at 37℃ for 1, 3, 5 and 7days respectively. Formation of cells in the sub-G1 phase was indicative of apoptotic cells. Cell viability and apoptosis rate were measured by the MTT assay, flow cytometry. Cells morphology were observed by transmission electron microscope. Results： Vincristine resistance in KBV cells was significantly higher than KB cells.The expression of IL-24 was detected in both transfected KB cells and KBV cells. IL-24 could inhibit the growth of KB cells and KBV cells, while no growth inhibition in Ha Ca T cells. Overexpression of IL-24 could induce apoptosis in KB and KBV cells, but there was no similar effect in Ha Ca T cells. Conclusion： Hybrid virus-mediated IL-24 had strong cytotoxicity and induction apoptosis effects on OSCC drug-resistant KBV cell line, and had no side effects on normal cells, which may develop a new way to treat oral drug resistant cancer.

关 键 词：IL-24 口腔鳞癌 耐药 凋亡 

分 类 号：R739.85[医药卫生—肿瘤]