中文摘要：

目的：探讨生物钟基因Period2对卵巢癌裸鼠移植瘤生长转移和血管生成的作用及可能机制。方法：利用卵巢癌细胞株构建裸鼠卵巢癌皮下移植瘤,利用基因转染技术,外源性导入重组基因Period2,使之在肿瘤组织成功稳定表达,分别采用Real-time定量PCR和Western blot检测移植瘤中Period2表达,治疗期间测量移植瘤体积,治疗2周后处死裸鼠,称取瘤重,免疫组化检测肿瘤组织中血管内皮生长因子/血管通透性因子（VEGF/VPF）、血管内皮生长因子受体1（VEGFR1）、微血管密度（MVD）（CD34标记）的表达情况,Western blot检测肿瘤转移相关基因（MTA1）、基质金属蛋白酶-9（MMP-9）,以及PI3K/Akt信号通路的表达。结果：（1）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。（2）Period2组移植瘤体积与其他两组相比较,差异有统计学意义（F=23.469,P〈0.001）。转染后2周,Period2组移植瘤重量明显低于空质粒组和对照组（P〈0.05）,Period2组抑瘤率达到38.9%。（3）免疫组化结果表明,Period2组的VEGF/VPF、VEGFR1表达下降（F=46.80/48.09,P〈0.001）,MVD计数（CD34标记）显著减少（F=138.4,P〈0.001）。（4）Western blot结果表明,Period2组MTA-1和MMP-9表达明显少于其他组（P〈0.05）,自噬与凋亡相关信号通路PI3K/Akt中标志性蛋白PI3K、Akt表达明显下调（P〈0.05）。结论：（1）外源性导入Period2过表达可使卵巢癌生长速度减慢,抑瘤率明显提高。（2）Period2可能通过抑制VEGF/VPF、MTA-1、MMP-9表达而抑制卵巢癌的血管新生和浸润转移。（3）Period2可能通过干扰PI3K/Akt信号通路影响凋亡,抑制肿瘤血管形成来发挥抑瘤作用。

英文摘要：

Objective： To investigate the effects of circadian gene Period2 on the growth and angiogenesis of ovarian cancer xenografts in nude mice. Methods： Using ovarian cancer cell to construct ovarian cancer xenografts in nude mice,the exogenous gene Period2 was introduced into the recombinant gene by gene transfer technique,which was successfully expressed in tumor tissue. Real-time quantitative PCR and Western blot were used to detect the expression of Period2 in xenografts tumors.The volume of xenografts tumors were measured during the treatment period,and after two weeks of treatment,the mice were sacrificed and the weight of the xenografts tumors were measured. The expression of vascular endothelial growth factor/vascular permeability factor VEGF/VPF,vascular endothelial growth factor receptor1（ VEGFR1） and microvessel density MVD（ CD34 marks） in tumor tissue were detected by im-munohistochemistry.The expression of tumor metastasis associated gene MTA1,matrix metalloproteinase MMP-9,and PI3K/Akt signaling pathway was detected by western-blot method. Results：（ 1） Successful expression of exogenous Period2 gene in transplanted tumor in nude mice.（ 2） Compared with the other two groups,the tumor volume of Period2 group was statistically significant（ F = 23.469,P0.001）.2 weeks after transfection,transplanted tumor weight of Period2 group was significantly lower than the empty plasmid group and the control group,the difference was statistically significant（ P〈0. 05）,and the tumor inhibition rate of Period2 reached 38.9%.（ 3） Immunohistochemical results showed that VEGF/VPF,VEGFR1 expression decreased（ F = 46.80/48.09,P〈0.001）,and MVD counts（ CD34 markers） were significantly reduced（ F = 138.4,P〈 0.001） in the Period2 group.（ 4） The results of Western blot showed that,MTA1 and MMP-9 of Period2 group were significantly less than that of the other groups（ P0.05）,and the protein expression of autophagy related signaling pathway PI3K/Akt was significantly reduced（