CD137L基因转染对小鼠体内卵巢癌细胞生长的影响及其免疫调节研究

Effect of CD137L gene transfection on growth of ovarian cancer cells and its immune regulation in vivo

目的:初步探讨转染CD137L基因的小鼠卵巢癌细胞株OVHM细胞体内的致瘤性及其对免疫功能的影响。方法:以脂质体为载体将重组质粒pcDNA3.1和pcD-NA3.1/CD137LL分别导入OVHM细胞中,经HygroB筛选,用RT-PCR法检测目的基因的表达,筛选建立高表达的细胞株。计数法绘制体外培养细胞生长曲线。将转染前后的肿瘤细胞,分别接种于B6C3F1小鼠的腹部皮下观察其致瘤性。LDH法测定其CTL细胞的杀伤活性。结果:与野生型OVHM细胞相比,所建立的OVHM/CD137L细胞株可高表达CD137L。转染前后肿瘤细胞的体外生长速度无明显变化。OVHM/CD137L细胞在小鼠体内的致瘤性较OVHM/pcDNA3.1细胞和野生型OVHM细胞明显降低。接种OVHM/CD137L细胞小鼠的CTL细胞杀伤活性明显增强。结论:转染CD137L基因后OVHM细胞在小鼠体内的致瘤性显著降低,且诱导产生了明显的抗肿瘤免疫。CD137L有望成为卵巢癌基因治疗的候选基因。

Objective：To observe the effect of CD137L gene transfeetion on mouse ovarian cancer cell growth and its immune regulation. Methods：Mouse ovarian cancer cell lines （OVHM） were transfected with pcDNA3. 1 and pcDNA3. 1/CD137L through LipofectamineTM2000 mediation. The expression of CD137L gene was detected by RT-PCR and the high expression cell strain was selected. The growth curve was drawn by counting the number of the tumor cells. The tumor size was observed after VHM/CD137L cells, OVHM/pcDNA3.1 cells and parental OVHM cells were injected subcutaneously into B6C3F1 mouse. The activity of CTL from the spleen of immunized mouse was detected with lactate dehydrogenase （LDH） method using parental OVHM cell as the target cells. Results：Compared with the other two groups, the expression of CD137L mRNA in OVHM/CD137L cells increased markedly. There was no difference between the proliferation rates of transfected and untransfected cells in vitro. Compared with the other two groups, delayed tumor formation and slow tumor growth were observed in mice immunized with OVHM/CD137L cells. There was no difference in tumor growth between VHM/CD137L group and OVHM/pcDNA3.1 group. The CTL activity in B6C3F1 spleen mice immunized by OVHM/CD137L cell was significantly enhanced. Conclusion：CD137L may be a candidate gene in immunogene therapy of ovarian cancer.