腺病毒介导HSV-TK/GCV系统治疗口腔鳞癌机制的研究

Therapeutic Mechanism of Adenovirus Mediated HSV-TK/GCV for Oral Squamous Carcinoma Cells in Vitro

【目的】研究腺病毒介导的单纯疱疹病毒胸苷激酶基因 (AdCMVHSV TK) /丙氧鸟苷 (GCV)系统治疗口腔鳞癌细胞的作用机制。【方法】HSV TK/GCV系统治疗口腔鳞癌细胞 (Tca 8113细胞株 )后分别应用3 H TdR掺入法、流式细胞仪(FCM )、TUNEL(TdT mediateddUTPNickEndLabeling)法和透射电镜等方法进行检测。【结果】HSV TK/GCV系统治疗后 :3 H TdR掺入法显示掺入率下降 ,GCV为 5× 10 -4 mol/L时掺入率随感染复数 (MOI)的上升而显著下降 (P <0 0 0 1) ;与对照组比较 ,FCM显示S期比率显著升高 (P <0 0 0 1) ,G2 +M期比率下降为 0 (P <0 0 0 1) ,G1/G0 期的改变不明显 (P>0 0 5 ) ;透射电镜及TUNEL法检测均未见凋亡细胞。【结论】口腔鳞癌细胞经HSV TK/GCV系统治疗后DNA的合成受抑制 ,细胞周期阻滞于 S期 ,但无明显诱导细胞凋亡作用。

To determine the therapeutic mechanism of adenovirus mediated herpes simplex virus thymidine kinase (AdCMV HSV TK)/ganciclovir (GCV) suicide gene therapy for oral squamous carcinoma cells (Tca 8113 cell line) in vitro. 3H thymidine( 3H TdR) incorporation assay, flow cytometry(FCM), transmission electron microscope and TUNEL(TdT mediated dUTP Nick End Labeling) assay were used to detected the changes of Tca 8113 cells after treated with AdCMV HSV TK/GCV system. After treated with AdCMV HSV TK/GCV system, the 3H TdR incorporation rate decreased and significant decreased between different multiple of infection (MOI) at GCV 5×10 -4 mol/L(P<0 001). Compared with control groups, the AdCMV HSV TK/GCV treated cells accumulated in S phase of cell cycle (P<0 001), G 2+M phase dropped to zero (P<0 001), and G 1/G 0 phase didn't change (P>0 05). Apoptotic cells can't be found in AdCMV HSV TK/GCV treated cells under transmission electron microscope and TUNEL assay. [Conclusion] The killing effects of HSV TK/GCV system on oral squamous carcinoma cells maybe inhibit DNA synthesis, block cell cycle in S phase, but can't induce apoptosis.