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目的:探讨生物钟基因Period2对卵巢癌裸鼠移植瘤生长抑制的作用机制。方法:采用卵巢癌SKVO3细胞株构建裸鼠卵巢癌移植瘤,利用基因转染技术外源性导入重组基因质粒Period2。采用Real-time PCR法和Western bolt法检测移植瘤中Period2表达,治疗期间测量移植瘤体积。治疗后2周处死裸鼠,称取瘤重,采用Real-time PCR和Western blot法检测肿瘤组织中Bcl-2和Bax基因的表达。结果:外源性导入Period2基因在裸鼠移植瘤肿瘤组织中成功稳定表达。与对照组比较,Period2基因过表达组的肿瘤生长速度减慢,肿瘤体积和重量降低,抑瘤率升高,凋亡基因Bax表达明显升高,凋亡抑制基因Bcl-2表达明显降低,差异均有统计学意义(P<0.05)。结论:Period2可能通过下调肿瘤凋亡抑制基因Bcl-2,促进凋亡基因Bax表达,促进肿瘤细胞凋亡,从而抑制卵巢癌的生长转移,发挥抑瘤作用。
Objective: To investigate the mechanism of biological clock gene Period2 on the growth inhibition of ovarian cancer xenografts in nude mice. Methods: The ovarian cancer xenografts in nude mice were constructed by ovarian cancer cell line SKVO3. The gene transfection technique was used to introduce the recombinant plasmid pE2 gene periodically. Real-time PCR and Western blot were used to detect the expression of Period2 in the xenografts and the volume of xenografts was measured during the treatment. The nude mice were sacrificed 2 weeks after the treatment, and the tumor weights were weighed. The expression of Bcl-2 and Bax gene in tumor tissues was detected by Real-time PCR and Western blot. Results: The exogenous introduction of Period2 gene was successfully and stably expressed in the xenografted tumor of nude mice. Compared with the control group, the growth of tumor cells in Period2 gene over-expression group was slowed down, tumor volume and weight were decreased, tumor inhibition rate was increased, apoptosis gene Bax expression was significantly increased, and apoptosis inhibitor gene Bcl-2 expression was significantly decreased All were statistically significant (P <0.05). Conclusion: Period2 may inhibit the growth and metastasis of ovarian cancer by inhibiting the expression of Bcl-2, decreasing the expression of Bax and promoting the apoptosis of tumor cells.