动脉灌注GE7系统介导1型单纯疱疹病毒胸腺嘧啶核苷激酶／羟甲基无环鸟苷对大鼠诱发性卵巢恶性肿瘤的作用

Therapeutic effect of ovarian intra-arteriai infusion of GET-defivery system-mediated HSVI-tk/ ganciciovir gene therapy in a rat model of malignant ovarian tumor

目的观察非病毒载体介导1型单纯疱疹病毒胸腺嘧啶核苷激酶（HSVl-tk）／羟甲基无环鸟苷（GCV）基因治疗系统，经肿瘤供血动脉灌注，对大鼠诱发性卵巢癌的作用，并初步探讨其作用机制。方法构建GE7-polylysine／pCMV-HSVl-tk／polylysine-HA20四元复合物，取二甲基苯蒽（DMBA）诱发的大鼠卵巢肿瘤模型18只，随机分为3组，每组6只。分别经卵巢供血动脉注入四元复合物8“g／只（Atk组）、同体积的生理盐水（ANS组），经尾静脉注入四元复合物8μg／只（Vtk组）。72h后，3组大鼠均腹腔注射GCV，50mg·kg-1.d-1，连用10d。停药后3d处死大鼠，瘤体称重，计算抑瘤率。采用流式细胞仪检测各组大鼠卵巢肿瘤细胞的细胞周期变化和细胞凋亡情况。结果Atk组大鼠肿瘤瘤重为（4．774-2．31）g，明显低于ANS组[（14．66±6．26）g，P〈0．01]和Vtk组[（17．53±7．19）g，P〈0．01]。Atk组抑瘤率为67．5％Vtk组的抑瘤率为-19．6％。流式细胞检测结果显示，Atk组S期细胞占（54．32±9．65）％，高于ANS组[（27．434-9．22）％，P〈0．01]和Vtk组[（30．16±11．57）％，P〈0．01]。Ark组肿瘤细胞凋亡率为（39．15±12．16）％，高于ANS组[（11．864-5．28）％，P〈0．01]和Vtk组[（14．32±6．43）％，P〈0．01]。结论HSVl-tk在GE7导人系统介导下，经卵巢供血动脉导人大鼠卵巢肿瘤组织，给予GCV后，可使肿瘤细胞阻滞于S期，促进肿瘤细胞凋亡，从而发挥良好的抑瘤作用。

Objective To observe the gene expression of herpes simplex virus type 1 thymidine kinase （HSVI-tk） in rat malignant ovarian tumor tissues and the therapeutic effect of ganciclovior （GCV） after intra-arterial infusion of HSVI-tk gene therapy mediated by GE7-delivery system. Methods A GET- polylysine/pCMV-HSVl-tk/polylysine-HA20 4-element complex was constructed. Eighteen rats with DMBAinduced ovarian tumor were divided into 3 groups as Atk, ANS and Vtk groups. The 4-element complex GE7-polylysine/pCMV-HSVl-tk/polylysine-HA20 was injected via the ovarian artery into the rats of Atk group, saline buffer was injected in the ANS groups, and the 4-element complex was injected via the tail vein into the rats of Vtk group. All rats received intraperitoneal injection of GCV in a dose of 50 mg/kg daily for 10 days. The rats were sacrificed 3 days after the final dose of GCV, and the tumor weight was measured and tumor growth inhibition rate was calculated. Flow cytometry was used to assess the cell cycle and apoptosis. Results The tumor weight in the rats of Atk group was （4.77 ~ 2.31 ） g, significantly lower than that of ANS group [（14.66±6.26）g, P〈0.01] and Vtk group [（17.53 ±7.19）g, P〈0.01]. The tumor growth inhibition rate of the Atk group was 67.5%, while that of Vtk group was - 19.6%. The flowcytometry showed that S-phase tumor cells in the Atk group were （ 54.32 ± 9.65 ） % , significantly higher than that in the ANS （27.43±9.22）% and （30.16 ±11.57）% in the Vtk group （both P〈0.01）. The tumor cell apoptosis rate in the Ark group was （39.15 ± 12.16）% , significantly higher than that in the ANS group [ （ 11.86 ± 5.28） %, P 〈 0.01 ] and Vtk group [ （ 14.32 ± 6.43） %, P 〈 0. 01 ]. Conclusion HSVI-tlc/ GCV gene therapy system mediated by GE7 non-viral delivery system via ovarian arterial infusion effectively causes cell cycle arrest at S phase and enhances cell apoptosis, therefore, exerts an inhibitory effect on tumor growth.