生物钟基因Period2对裸鼠卵巢癌移植瘤生长和血管生成抑制作用的机制研究

发布时间：2018-06-13 18:34

本文选题：卵巢癌 + 生物钟基因Period　；参考：《现代妇产科进展》2017年03期

【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,P0.001)。转染后2周,Period2组移植瘤重量明显低于空质粒组和对照组(P0.05),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,P0.001),MVD计数(CD34标记)显著减少(F=138.4,P0.001)。(4)Western blot结果表明,Period2组MTA-1和MMP-9表达明显少于其他组(P0.05),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(P0.05)。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。
[Abstract]:Aim: to investigate the effect and possible mechanism of biological clock gene Period2 on growth and angiogenesis of ovarian cancer xenografts in nude mice. Methods: nude mice ovarian cancer subcutaneous transplanted tumor was constructed by using ovarian cancer cell line. The recombinant gene Period2 was introduced into the tumor tissue successfully and stably by gene transfection technique. Real-time quantitative PCR and Western blot were used to detect the expression of Period2 in the transplanted tumor. The volume of the transplanted tumor was measured during the treatment. After 2 weeks of treatment, the nude mice were killed and the tumor was weighed. Expression of vascular endothelial growth factor (VEGF) / vascular permeability factor (VEGF), vascular endothelial growth factor receptor (VEGF), vascular endothelial growth factor receptor (VEGFR1) and microvessel density (MVD) in tumor tissues were detected by immunohistochemistry. Western blot was used to detect the tumor metastasis related gene (MTA1) and matrix metal (MTA1). The expression of MMP-9 and PI3 K / Akt signaling pathway. Results compared with the other two groups, the volume of Period2 gene successfully expressed stably in xenograft tumor tissue of nude mice was significantly higher than that of the other two groups (P 0.001). 2 weeks after transfection, the weight of transplanted tumor in Period2 group was significantly lower than that in blank plasmid group and control group (38.9%). The immunohistochemical results showed that the expression of VEGF / VPFFR1 decreased in Period2 group (46.80 / 48.09P0.001MVD / CD34). The results of blot showed that MTA-1 and MMP-9 in Period2 group were significantly reduced. The expression of PI3K-1 Akt in autophagy and apoptosis-related signaling pathway (PI3K / Akt) was significantly lower than that in other groups (P 0.05). Conclusion the overexpression of Period2 can slow down the growth rate of ovarian cancer. The inhibition rate of Period2 may inhibit the angiogenesis, invasion and metastasis of ovarian cancer by inhibiting the expression of VEGF / VPFN MTA-1MTA-1MTA-1MMP-9, which may influence apoptosis by interfering with the PI3K / Akt signal pathway. Inhibition of tumor angiogenesis to play a role in tumor inhibition.
【作者单位】：山西医科大学第一医院妇科;
【基金】：山西省卫生计生委科研课题(No:2015024)
【分类号】：R737.31

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