發(fā)布時(shí)間：2018-06-13 18:34

  本文選題：卵巢癌 + 生物鐘基因Period　； 參考：《現(xiàn)代婦產(chǎn)科進(jìn)展》2017年03期

【摘要】：目的:探討生物鐘基因Period2對(duì)卵巢癌裸鼠移植瘤生長(zhǎng)轉(zhuǎn)移和血管生成的作用及可能機(jī)制。方法:利用卵巢癌細(xì)胞株構(gòu)建裸鼠卵巢癌皮下移植瘤,利用基因轉(zhuǎn)染技術(shù),外源性導(dǎo)入重組基因Period2,使之在腫瘤組織成功穩(wěn)定表達(dá),分別采用Real-time定量PCR和Western blot檢測(cè)移植瘤中Period2表達(dá),治療期間測(cè)量移植瘤體積,治療2周后處死裸鼠,稱(chēng)取瘤重,免疫組化檢測(cè)腫瘤組織中血管內(nèi)皮生長(zhǎng)因子/血管通透性因子(VEGF/VPF)、血管內(nèi)皮生長(zhǎng)因子受體1(VEGFR1)、微血管密度(MVD)(CD34標(biāo)記)的表達(dá)情況,Western blot檢測(cè)腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)、基質(zhì)金屬蛋白酶-9(MMP-9),以及PI3K/Akt信號(hào)通路的表達(dá)。結(jié)果:(1)外源性導(dǎo)入Period2基因在裸鼠移植瘤腫瘤組織中成功穩(wěn)定表達(dá)。(2)Period2組移植瘤體積與其他兩組相比較,差異有統(tǒng)計(jì)學(xué)意義(F=23.469,P0.001)。轉(zhuǎn)染后2周,Period2組移植瘤重量明顯低于空質(zhì)粒組和對(duì)照組(P0.05),Period2組抑瘤率達(dá)到38.9%。(3)免疫組化結(jié)果表明,Period2組的VEGF/VPF、VEGFR1表達(dá)下降(F=46.80/48.09,P0.001),MVD計(jì)數(shù)(CD34標(biāo)記)顯著減少(F=138.4,P0.001)。(4)Western blot結(jié)果表明,Period2組MTA-1和MMP-9表達(dá)明顯少于其他組(P0.05),自噬與凋亡相關(guān)信號(hào)通路PI3K/Akt中標(biāo)志性蛋白PI3K、Akt表達(dá)明顯下調(diào)(P0.05)。結(jié)論:(1)外源性導(dǎo)入Period2過(guò)表達(dá)可使卵巢癌生長(zhǎng)速度減慢,抑瘤率明顯提高。(2)Period2可能通過(guò)抑制VEGF/VPF、MTA-1、MMP-9表達(dá)而抑制卵巢癌的血管新生和浸潤(rùn)轉(zhuǎn)移。(3)Period2可能通過(guò)干擾PI3K/Akt信號(hào)通路影響凋亡,抑制腫瘤血管形成來(lái)發(fā)揮抑瘤作用。
[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.
【作者單位】： 山西醫(yī)科大學(xué)第一醫(yī)院婦科;
【基金】：山西省衛(wèi)生計(jì)生委科研課題(No:2015024)
【分類(lèi)號(hào)】：R737.31

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本文編號(hào)：2015005