本文關(guān)鍵詞： 子宮內(nèi)膜癌 人附睪蛋白 侵襲 轉(zhuǎn)染 實(shí)時定量聚合酶鏈反應(yīng) 人　出處：《解剖學(xué)報》2017年06期 　論文類型：期刊論文

【摘要】：目的探討人附睪蛋白4(HE4)過表達(dá)對子宮內(nèi)膜癌(EC)細(xì)胞增殖、侵襲能力及腫瘤形成的影響。方法構(gòu)建HE4過表達(dá)質(zhì)粒載體(pc DNA 3.1/Myc-His-HE4)和HE4特異性小干擾RNA(siRNA)表達(dá)質(zhì)粒載體(siRNA-HE4),分別轉(zhuǎn)染HEC-1B(HE4過表達(dá)組)和Ark2(HE4低表達(dá)組)兩個EC細(xì)胞系;以空載體pc DNA 3.1/Myc-His轉(zhuǎn)染的HEC-1B細(xì)胞為HE4過表達(dá)組的陰性對照,以非特異性序列siRNA轉(zhuǎn)染的Ark2細(xì)胞為HE4低表達(dá)組的陰性對照;不進(jìn)行轉(zhuǎn)染處理、正常培養(yǎng)的EC細(xì)胞系為正常對照組。轉(zhuǎn)染后,采用實(shí)時定量聚合酶鏈反應(yīng)(Real-time PCR)法檢測HE4 mRNA表達(dá)水平,MTT比色法測定細(xì)胞增殖活性,體外遷移和侵襲實(shí)驗(yàn)檢測細(xì)胞的遷移和侵襲能力。構(gòu)建移植性腫瘤小鼠模型,比較各組動物的形成瘤體積。結(jié)果與正常對照組和陰性對照組的HEC-1B細(xì)胞比,轉(zhuǎn)染pc DNA 3.1/Myc-His-HE4后HEC-1B細(xì)胞中HE4 mRNA表達(dá)水平顯著升高(P0.05),細(xì)胞增殖活性、穿膜細(xì)胞數(shù)明顯升高(P0.05);與正常對照組和陰性對照組的Ark2細(xì)胞比,轉(zhuǎn)染siRNA-HE4后Ark2細(xì)胞中HE4 mRNA表達(dá)水平顯著降低(P0.05),細(xì)胞增殖活性、穿膜細(xì)胞數(shù)明顯降低(P0.05);而陰性對照組與正常對照組之間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05);HE4過表達(dá)組的移植性腫瘤體積和重量明顯大于其對照組(P0.05),而HE4低表達(dá)組的移植性腫瘤體積和重量明顯小于其對照組(P0.05)。結(jié)論 HE4過表達(dá)可導(dǎo)致EC細(xì)胞的增殖、遷移及侵襲能力增強(qiáng),促進(jìn)腫瘤形成;下調(diào)HE4基因表達(dá)可明顯抑制EC細(xì)胞的增殖、遷移和侵襲,并抑制腫瘤生長。
[Abstract]:Objective to investigate the effect of overexpression of human epididymal protein 4 (HE4) on the proliferation of endometrial carcinoma (EC) cells. Methods the overexpression plasmid of HE4 was constructed to construct the pc DNA 3.1% Myc-His-HE4 and HE4 specific small interference RNAs. SiRNA-HE4). Two EC cell lines were transfected into HEC-1B(HE4 overexpression group and Ark2(HE4 low expression group, respectively. HEC-1B cells transfected with empty vector PC DNA 3.1% Myc-His were used as negative control in HE4 overexpression group. Ark2 cells transfected with nonspecific sequence siRNA were used as negative control in HE4 low expression group. Without transfection treatment, the normal cultured EC cell line was the normal control group. The expression level of HE4 mRNA was detected by real-time quantitative polymerase chain reaction real-time PCR. In vitro migration and invasion experiments were used to detect the migration and invasion ability of cells. The volume of tumor formation in each group was compared with that of normal control group and negative control group. The results were compared with those of normal control group and negative control group. After transfection of PC DNA 3.1% Myc-His-HE4, the expression of HE4 mRNA in HEC-1B cells increased significantly. The number of perforating cells increased significantly (P 0.05). Compared with the normal control group and the negative control group, the expression of HE4 mRNA in Ark2 cells after siRNA-HE4 transfection was significantly lower than that in the normal control group and negative control group (P 0.05). The cell proliferation activity and the number of transmembrane cells decreased significantly (P 0.05). There was no significant difference between the negative control group and the normal control group (P 0.05). The volume and weight of transplanted tumor in HE4 overexpression group was significantly larger than that in control group (P0.05). However, the volume and weight of transplanted tumor in the low expression group of HE4 were significantly smaller than that in the control group (P 0.05). Conclusion overexpression of HE4 can increase the proliferation, migration and invasion of EC cells. Promoting tumor formation; Down-regulation of HE4 gene expression can significantly inhibit the proliferation, migration and invasion of EC cells, and inhibit tumor growth.
【作者單位】： 承德醫(yī)學(xué)院附屬醫(yī)院婦科;承德醫(yī)學(xué)院附屬醫(yī)院產(chǎn)科;承德醫(yī)學(xué)院附屬醫(yī)院檢驗(yàn)科;
【基金】：河北省人口和計(jì)劃生育委員會科技研究計(jì)劃項(xiàng)目(2012-A25) 承德市科技支撐計(jì)劃項(xiàng)目(2013-2044)
【分類號】：R737.33
【正文快照】： 子宮內(nèi)膜癌(endometrial cancer,EC)是發(fā)生于子宮內(nèi)膜的一組上皮惡性腫瘤,為女性3大生殖系統(tǒng)惡性腫瘤之一,具有較高的發(fā)病率和死亡率。前期臨床研究表明,近年發(fā)現(xiàn)的新型腫瘤標(biāo)志物人附睪蛋白4(human epididymal protein 4,HE4)對EC的診斷、疾病進(jìn)展及預(yù)后評估中具有重要作用[1

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