《口腔颌面外科杂志》 ›› 2015, Vol. 25 ›› Issue (4): 246-. doi: 10.3969/j.issn.1005-4979.2015.04.002

• 基础研究 • 上一篇    下一篇

病毒介导IL-24基因对口腔鳞癌耐药细胞株凋亡的影响

杨德钊1,王丹丹2,孟祥1,杨帅1,孙宏晨2,李吉辰1   

  1. 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
  • 出版日期:2015-08-28 发布日期:2015-12-01
  • 通讯作者: 李吉辰,教授. E-mail:lijichen@163.com
  • 作者简介:杨德钊(1989—),男,黑龙江省牡丹江市人,硕士研究生. E-mail: 264706737@163.com
  • 基金资助:

    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)

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

YANG De-zhao1, WANG Dan-dan2, MENG Xiang1, YANG Shuai1, SUN Hong-chen2, LI Ji-chen1   

  1. 1. Department of Oral and Maxillofacial Surgery, Harbin Medical University Hospital of Stomatology, Harbin 150001, Heilongjiang Province;2. Department of Pathology, School of Stomatology, Jilin University, Changchun 130021, Jilin Province, China
  • Online:2015-08-28 Published:2015-12-01

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AdLTR2EF1α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HaCaT细胞作为对照,使用AdLTR2EF1α-IL-24转染KB、KBV及HaCaT细胞后,MMT法检测细胞活力, AnnexinV/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。AdLTR2EF1α-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HaCaT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。

关键词: IL-24;  , 口腔鳞癌;  , 耐药;  , 凋亡

Abstract: 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, AdLTR2EF1α-EGFP or AdLTR2EF1α-IL-24 were used to treat the KB cells in order to obtain the optimal drug concentration. Human keratinocyte HaCat cells were used as control cells. AdLTR2EF1α-IL-24 and AdLTR2EF1α-vec transfected the KB ,KBV, and HaCat 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 HaCaT cells. Overexpression of IL-24 could induce apoptosis in KB and KBV cells, but there was no similar effect in HaCaT 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.

Key words: interleukin-24, oral squamous cell carcinoma, drug resistance, apoptosis

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