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- 2017
IL-18基因联合二萜生物碱对舌鳞癌细胞的增殖抑制与促凋亡作用
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Abstract:
摘要：目的 探讨人IL-18基因联合二萜生物碱对舌鳞癌细胞Tscca的增殖抑制与促凋亡作用。方法 构建pEGFPN3-IL-18质粒，转染舌鳞癌细胞Tscca。荧光显微镜下观察质粒的转染效率；MTT检测pEGFPN3-IL-18联合二萜生物碱对舌鳞癌细胞Tscca生长的影响；流式细胞术检测细胞凋亡情况；蛋白质免疫印迹法检测细胞信号调节激酶Akt/p-Akt的蛋白水平。结果 转染pEGFPN3-IL-18的舌鳞癌细胞Tscca较未转染细胞凋亡数增加，差异有统计学意义(P<0.05)；0.2、0.4、0.6mg/mL二萜生物碱培养舌鳞癌细胞，随浓度增加，细胞凋亡数增多，差异有统计学意义(P<0.05)；IL-18与二萜生物碱联合对Tscca细胞作用48h后，较单独使用二萜生物碱的增殖抑制作用明显，且呈浓度依赖性，差异有统计学意义(P<0.05)；IL-18联合二萜生物碱还可剂量依赖性地降低 p-Akt的蛋白水平。结论 pEGFPN3-IL-18联合二萜生物碱对舌鳞癌细胞Tscca有协同抑制作用。
ABSTRACT: Objective To investigate the effects of human IL-18 gene combined with diterpenoid alkaloids in inhibiting the proliferation and inducing the apoptosis of tongue squamous carcinoma cells Tscca. Methods We constructed recombinant plasmid pEGFPN3-IL-18 and tranfected it into tongue squamous carcinoma cells Tscca. The transduction efficiency of the target cells was detected by fluorescent microscopy, cytotoxic effect of IL-18 gene with diterpenoid alkaloids on Tscca was detected by MTT assay, and apoptosis was detected by flow cytometry. Western blot was employed to examine the expression level of cellular signal-regulated kinase Akt/p-Akt. Results The tongue squamous cells Tscca which transfected pEGFPN3-IL-18 had significantly increased apoptosis compared with non-transfected cells (P<0.05). Tongue carcinoma squamous cells cultured with diterpenoid alkaloids at the concentrations of 0.2, 0.4 and 0.6mg/mL had significantly increased apoptosis in a dose-dependent manner (P<0.05). Human IL-18 gene combined with diterpenoid alkaloids for 48 hours inhibited significantly Tscca in a concentration-dependent manner compared with diterpenoid alkaloids alone (P<0.05). The two in combination could also decrease the protein level of p-Akt dose-dependently. Conclusion The combination of pEGFPN3-IL-18 and diterpenoid alkaloids has a synergistic effect in inhibiting the growth of tongue squamous carcinoma cells Tscca
| [1] | 梁欣娜，张兴？觯？滕红丽. 中药及其有效成分抗肿瘤作用研究[J].时珍国医国药, 2013, 24(1):119-122. |
| [2] | SUN GW, TANG EY, YANG XD, et al. Salvage treatment for recurrent oral squamous cell carcinoma[J]. J Craniofac Surg, 2009, 20(4):1093-1096. |
| [3] | CHODOEVA A, BOSC JJ, GUILLON J, et al. 8-O-Azeloyl-14-benzoylaconine: a new alkaloid from the roots of Aconitum karacolicum Rapcs and its antiproliferative activities[J]. Bioorg Med Chem, 2005, 13(23):6493-6501. |
| [4] | 徐秋萍，刘静涵，刘宝瑞. C19-、C20-二萜生物碱抗肿瘤活性研究进展[J]. 药学进展, 2016, 40(1):3-10. |
| [5] | 刘芳，杨春华，梁敬钰，等. C19型二萜生物碱药理活性和毒性研究进展[J]. 海峡药学, 2012, 24(4):1-5. |
| [6] | 彭勇泉，蔡捷，苏承武. 舌癌基因治疗的研究进展[J]. 蛇志, 2007, 19(4):290-293. |
| [7] | WIGGINTON JM, LEE JK, WILTROUT TA, et al. Synergistic engagement of ineffective endogenous anti-tumor immune response and induction of IFN-gamma and Fas-ligand-dependent tumor eradication by combined administration of IL-18 and IL-2[J]. J Immunol, 2002, 169(8):4467-4474. |
| [8] | 李峨，陈信义. 乌头碱抗耐药性人口腔鳞状上皮癌细胞作用的体外研究[J]. 中国中医药信息杂志, 2004, 11(2):103-105. |
| [9] | WANG CQ, LI YJ, WEI ZM, et al. Stable gene-silence of Kif2a synergistic with 5-fluorouracil suppresses oral tongue squamous cell carcinoma growth in vitro and in vivo[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2013, 116(1):49-54. |
| [10] | 刘炜炜，高尚，王战鑫，等. IL-18转染人舌癌CRL1623细胞及抗瘤作用的初步探讨[J]. 口腔医学研究, 2013, 29(3):233-237. |
| [11] | 李瑞阳，冯锋，刘静涵. C20型二萜生物碱的构效关系研究进展[J]. 海峡药学, 2013, 25(12):1-4. |
| [12] | WADA K, HAZAWA M, TAKAHASHI K, et al, Structure-activity relationships and the cytotoxic effects of novel diterpenoid alkaloid derivatives against A549 human lung carcinoma cells[J]. J Nat Med, 2011, 65(1):43-49. |
| [13] | SHAO Y, ZHANG SQ, QUAN F, et al. MicroRNA-145 inhibits the proliferation, migration and invasion of the human TCA8113 oral cancer line[J]. Oncol Lett, 2013, 6(6):1636-1640. |
| [14] | 倪晓辰，刘爽，张爱莉，等. IL-18基因转染人膀胱癌T-24细胞及其表达的研究[J]. 中国肿瘤临床, 2008, 35(11):652-655. |
| [15] | DE IN？IS C, REINA M, GAV？PN JA, et al. In vitro cytotoxicity of norditerpenoid alkaloids[J]. Z Naturforsch C, 2006, 61(1/2):11-18. |
| [16] | 田劭丹，刘雪强，王笑民，等. 乌头碱影响KBV<sub>200</sub>细胞Pgp蛋白表达的组化实验[J]. 中医药学刊, 2006, 24(1):55-56. |
| [17] | 张丹丹，李庆林. PI3K/Akt/mTOR信号通路与肿瘤[J]. 安徽医药，2012, 16(3): 281-283. |
| [18] | 刘田田，毕经旺. PI3K/Akt/mTOR信号通路与结直肠恶性肿瘤[J].实用医药杂志, 2016, 33(4):367-370. |
