發(fā)布時(shí)間：2018-05-30 12:19

  本文選題：子宮內(nèi)膜癌 + lncRNA��； 參考：《河北醫(yī)科大學(xué)》2017年博士論文

【摘要】：子宮內(nèi)膜癌是最常見(jiàn)的婦科惡性腫瘤之一。2015年子宮內(nèi)膜癌在中國(guó)的新發(fā)病例數(shù)約為63400人,死亡率約為21.8%,新發(fā)病例數(shù)逐年上升,且發(fā)病呈年輕化的趨勢(shì)。2016年美國(guó)子宮內(nèi)膜癌新發(fā)病例約為60050人,成為繼乳腺癌之后的第二大婦科癌癥,嚴(yán)重危害了女性的健康。盡管目前醫(yī)學(xué)診斷和治療水平已經(jīng)非常發(fā)達(dá),但子宮內(nèi)膜癌的確切發(fā)病機(jī)制尚不明確,治療手段也十分有限。最近,長(zhǎng)鏈非編碼RNA(long non-coding RNA,lncRNA)的發(fā)現(xiàn)為探索子宮內(nèi)膜癌的發(fā)病機(jī)制提供了分子生物學(xué)基礎(chǔ)。lncRNA是長(zhǎng)度超過(guò)200nt不能編碼蛋白質(zhì)的轉(zhuǎn)錄本,大量的研究發(fā)現(xiàn)其在人類(lèi)的生命活動(dòng)及各種疾病的發(fā)生發(fā)展中發(fā)揮了重要的作用。CARLo-5是最近發(fā)現(xiàn)的長(zhǎng)度為2628個(gè)核苷酸的lncRNA,位于原癌基因myc的增強(qiáng)子附近,據(jù)報(bào)道,CARLo-5在結(jié)腸癌、胃癌等多種癌組織中表達(dá)上調(diào),在癌癥的發(fā)生和進(jìn)展中起著重要作用,對(duì)多種癌癥的診斷、預(yù)后評(píng)估和治療均具有潛在價(jià)值。然而,CARLo-5在子宮內(nèi)膜癌中的表達(dá)情況及作用機(jī)制尚未見(jiàn)報(bào)道。本研究采用實(shí)時(shí)熒光定量PCR(Real-time PCR)的方法檢測(cè)正常子宮內(nèi)膜組織和子宮內(nèi)膜癌組織中CARLo-5基因的表達(dá)水平,使用蛋白印跡法(Western-blot)檢測(cè)細(xì)胞周期相關(guān)蛋白CDK2和CDKN1A的表達(dá)情況,深入分析了CARLo-5表達(dá)水平與子宮內(nèi)膜癌患者臨床病理特征和總生存期的相關(guān)性,并統(tǒng)計(jì)分析了CARLo-5基因表達(dá)水平與細(xì)胞周期相關(guān)蛋白CDK2和CDKN1A表達(dá)水平的相關(guān)性。體外敲低子宮內(nèi)膜癌細(xì)胞中CARLo-5的表達(dá)后,通過(guò)MTS法、Transwell遷移、Matrigel侵襲實(shí)驗(yàn)、平板克隆形成實(shí)驗(yàn)及流式細(xì)胞術(shù)檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞生物學(xué)行為的影響。為了探究CARLo-5在子宮內(nèi)膜癌中的作用機(jī)制,本研究通過(guò)Western-blot檢測(cè)了敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞中CDK2、CDKN1A、基質(zhì)金屬蛋白酶MMP2和MMP9表達(dá)水平的影響。通過(guò)RNAi慢病毒技術(shù)構(gòu)建了穩(wěn)定敲低CARLo-5的子宮內(nèi)膜癌細(xì)胞株,并進(jìn)行裸鼠皮下荷子宮內(nèi)膜癌細(xì)胞成瘤實(shí)驗(yàn),檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體內(nèi)增殖能力的影響。本研究探索了CARLo-5在子宮內(nèi)膜癌中的作用及機(jī)制,利于闡明子宮內(nèi)膜癌發(fā)生發(fā)展的分子生物學(xué)機(jī)制,從而為子宮內(nèi)膜癌的基因診斷和治療提供前期實(shí)驗(yàn)基礎(chǔ)。第一部分CARLo-5在子宮內(nèi)膜癌組織中的表達(dá)及臨床意義目的:研究lncRNA CARLo-5在子宮內(nèi)膜癌組織中的表達(dá)情況,并探討CARLo-5表達(dá)水平與子宮內(nèi)膜癌患者臨床病理特征及總生存期的關(guān)系。檢測(cè)子宮內(nèi)膜癌組織中細(xì)胞周期相關(guān)蛋白CDK2和CDKN1A的表達(dá)水平及其與CARLo-5表達(dá)水平的相關(guān)性。方法:1采用Real-time PCR法檢測(cè)108例子宮內(nèi)膜癌組織和66例正常子宮內(nèi)膜組織中CARLo-5的表達(dá)水平。2采用Western-blot檢測(cè)子宮內(nèi)膜癌組織和正常子宮內(nèi)膜組織中細(xì)胞周期相關(guān)蛋白CDK2和CDKN1A的表達(dá)情況。3分析CARLo-5的表達(dá)水平與子宮內(nèi)膜癌患者臨床病理特征和總生存期的相關(guān)性。4分析CARLo-5在子宮內(nèi)膜癌組織中的表達(dá)與細(xì)胞周期相關(guān)蛋白CDK2和CDKN1A表達(dá)之間的相關(guān)性。結(jié)果:1 lncRNA CARLo-5在子宮內(nèi)膜癌組織和正常子宮內(nèi)膜組織中的表達(dá)水平Real-time PCR實(shí)驗(yàn)結(jié)果顯示:CARLo-5在子宮內(nèi)膜癌中的表達(dá)水平較正常子宮內(nèi)膜組織明顯升高(P0.001)。2 CARLo-5的表達(dá)水平與子宮內(nèi)膜癌患者臨床病理特征的相關(guān)性CARLo-5在子宮內(nèi)膜癌組織中的表達(dá)水平與子宮內(nèi)膜癌患者的臨床分期和淋巴結(jié)轉(zhuǎn)移密切相關(guān)(P0.05),與患者的年齡、病理類(lèi)型、病理分級(jí)、肌層侵犯深度和脈管浸潤(rùn)無(wú)明顯相關(guān)性(P0.05)。3 CARLo-5的表達(dá)水平與子宮內(nèi)膜癌患者總生存期的相關(guān)性CARLo-5高表達(dá)組子宮內(nèi)膜癌患者的平均生存時(shí)間(43.0±14.1月)較CARLo-5低表達(dá)組患者(52.5±9.1月)明顯縮短(P0.05)。4子宮內(nèi)膜癌組織中CDK2和CDKN1A蛋白的表達(dá)水平Western-blot結(jié)果顯示:CDK2蛋白在子宮內(nèi)膜癌組織中的表達(dá)水平(9.32±0.69)較正常子宮內(nèi)膜組織(1.00±0.54)明顯升高,而CDKN1A蛋白在子宮內(nèi)膜癌組織中的表達(dá)水平(0.71±0.14)較正常子宮內(nèi)膜組織(1.00±0.11)明顯降低(P0.001)。5 CARLo-5的表達(dá)水平與CDK2和CDKN1A蛋白表達(dá)水平的相關(guān)性子宮內(nèi)膜癌組織中CDK2蛋白的表達(dá)水平與CARLo-5的表達(dá)呈正相關(guān)關(guān)系(r=0.30,P=0.043)。CDKN1A蛋白的表達(dá)與CARLo-5的表達(dá)呈負(fù)相關(guān)關(guān)系(r=-0.28,P=0.046)。結(jié)論:1 CARLo-5在子宮內(nèi)膜癌組織中的表達(dá)較正常子宮內(nèi)膜明顯升高,提示CARLo-5可能在子宮內(nèi)膜癌發(fā)生發(fā)展中發(fā)揮了促進(jìn)作用。2 CARLo-5高表達(dá)與子宮內(nèi)膜癌患者的臨床分期、淋巴結(jié)轉(zhuǎn)移和不良預(yù)后相關(guān),提示CARLo-5可能成為子宮內(nèi)膜癌判斷預(yù)后的分子標(biāo)志物。3 CDK2蛋白在子宮內(nèi)膜癌組織中的表達(dá)水平較正常子宮內(nèi)膜明顯升高,CDKN1A蛋白表達(dá)水平明顯降低,且CDKN1A和CDK2蛋白的表達(dá)與CARLo-5的表達(dá)水平具有明顯相關(guān)性。這些結(jié)果提示,CARLo-5可能部分通過(guò)調(diào)節(jié)CDK2和CDKN1A蛋白的表達(dá)而在子宮內(nèi)膜癌中發(fā)揮促癌作用。第二部分CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞生物學(xué)行為的影響及機(jī)制研究目的:通過(guò)調(diào)節(jié)CARLo-5在子宮內(nèi)膜癌細(xì)胞中的表達(dá)情況來(lái)探討其對(duì)子宮內(nèi)膜癌細(xì)胞生物學(xué)行為的影響及其可能的機(jī)制。方法:1采用MTS法檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體外增殖活性的影響。2采用Transwell遷移、劃痕愈合實(shí)驗(yàn)和Matrigel侵襲實(shí)驗(yàn)檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體外遷移和侵襲能力的影響。3采用平板克隆形成實(shí)驗(yàn)檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞克隆形成能力的影響。4采用流式細(xì)胞術(shù)檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞增殖周期的影響。5采用Real-time PCR和Western-blot檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞中CDK2和CDKN1A基因及蛋白表達(dá)水平的影響。6采用Western-blot檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞中MMP2和MMP9蛋白表達(dá)水平的影響。結(jié)果:1敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體外增殖活性的影響MTS法檢測(cè)結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞HEC-1-B和KLE的體外增殖活性明顯降低(P0.05)。2敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體外遷移和侵襲能力的影響Transwell遷移和劃痕愈合實(shí)驗(yàn)結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞HEC-1-B和KLE的體外遷移能力明顯降低(P0.05);Matrigel侵襲實(shí)驗(yàn)結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞HEC-1-B和KLE的體外侵襲能力明顯降低(P0.05)。3敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞克隆形成能力的影響平板克隆形成實(shí)驗(yàn)結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞HEC-1-B和KLE的克隆形成能力明顯下降(P0.05)。4敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞增殖周期的影響流式細(xì)胞術(shù)結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌HEC-1-B和KLE細(xì)胞G0/G1期細(xì)胞數(shù)明顯增多,S期細(xì)胞數(shù)明顯減少(P0.05)。5敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞CDK2和CDKN1A的基因及蛋白表達(dá)水平的影響Real-time PCR和Western-blot結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞中CDK2 m RNA和蛋白的表達(dá)水平明顯降低(P0.05),而CDKN1A m RNA和蛋白的表達(dá)水平明顯增高(P0.05)。6敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞MMP2和MMP9蛋白表達(dá)水平的影響Western-blot結(jié)果顯示:敲低CARLo-5后子宮內(nèi)膜癌細(xì)胞中MMP2和MMP9蛋白表達(dá)水平明顯降低(P0.05)。結(jié)論:1敲低CARLo-5能抑制子宮內(nèi)膜癌細(xì)胞的體外增殖、遷移和侵襲能力,提示CARLo-5的表達(dá)能夠促進(jìn)子宮內(nèi)膜癌細(xì)胞的增殖、侵襲和轉(zhuǎn)移,CARLo-5有望成為子宮內(nèi)膜癌治療的新的生物學(xué)靶標(biāo)。2敲低CARLo-5將子宮內(nèi)膜癌細(xì)胞阻滯在G0/G1期,并抑制CDK2 m RNA和蛋白水平的表達(dá),促進(jìn)CDKN1A m RNA和蛋白水平的表達(dá),提示CARLo-5可能通過(guò)調(diào)控CDK2和CDKN1A的表達(dá),抑制細(xì)胞G0/G1期阻滯,進(jìn)而促進(jìn)子宮內(nèi)膜癌細(xì)胞的增殖。3敲低CARLo-5能夠抑制子宮內(nèi)膜癌細(xì)胞中MMP2和MMP9蛋白的表達(dá),提示CARLo-5可能通過(guò)上調(diào)MMP2和MMP9蛋白的表達(dá)來(lái)促進(jìn)子宮內(nèi)膜癌的轉(zhuǎn)移。第三部分敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞裸鼠移植瘤生長(zhǎng)的影響及機(jī)制研究目的:檢測(cè)穩(wěn)定敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞在裸鼠體內(nèi)的成瘤能力的影響,通過(guò)裸鼠體內(nèi)實(shí)驗(yàn)來(lái)探討其在子宮內(nèi)膜癌發(fā)生發(fā)展中的作用機(jī)制。方法:1通過(guò)RNAi慢病毒技術(shù)構(gòu)建穩(wěn)定敲低CARLo-5的子宮內(nèi)膜癌細(xì)胞株。2裸鼠皮下成瘤實(shí)驗(yàn)檢測(cè)敲低CARLo-5對(duì)子宮內(nèi)膜癌細(xì)胞體內(nèi)成瘤能力的影響。3免疫組織化學(xué)法實(shí)驗(yàn)檢測(cè)裸鼠移植瘤組織中CDK2和CDKN1A蛋白的表達(dá)水平。結(jié)果:1敲低CARLo-5在體內(nèi)對(duì)子宮內(nèi)膜癌細(xì)胞的影響sh RNA-1組、sh RNA-2組和對(duì)照組裸鼠移植瘤的平均體積分別為(157.66±37.93 mm3)、(102.81±58.33mm3)、(499.74±98.61 mm3),敲低CARLo-5組裸鼠移植瘤的生長(zhǎng)速度和重量較未敲低組均明顯降低(P0.05)。2敲低CARLo-5對(duì)裸鼠移植瘤組織中CDK2和CDKN1A蛋白的表達(dá)水平的影響免疫組織化學(xué)結(jié)果顯示:敲低CARLo-5組裸鼠移植瘤組織中CDK2的蛋白表達(dá)水平較未敲低組明顯降低(P0.05),而CDKN1A的蛋白表達(dá)水平較未敲低組明顯增高(P0.05)。結(jié)論:敲低CARLo-5明顯抑制子宮內(nèi)膜癌細(xì)胞的體內(nèi)增殖活性,其作用機(jī)制可能為CARLo-5抑制CDKN1A并促進(jìn)CDK2蛋白表達(dá),進(jìn)而促進(jìn)子宮內(nèi)膜癌細(xì)胞的體內(nèi)增殖,CARLo-5有望成為子宮內(nèi)膜癌治療的新的靶點(diǎn)。
[Abstract]:Endometrial carcinoma is one of the most common gynecologic malignancies in.2015 years. The number of new cases of endometrial cancer in China is about 63400, the mortality rate is about 21.8%. The number of new cases is rising year by year, and the incidence is younger in.2016. The new incidence of endometrial cancer in the United States is about 60050, and the second major gynecologic cancer after breast cancer. In spite of the serious harm to the health of women, although the level of medical diagnosis and treatment is very advanced, the exact pathogenesis of endometrial cancer is not clear and the treatment is very limited. Recently, the discovery of long chain non coded RNA (long non-coding RNA, lncRNA) provides molecular biology for the pathogenesis of endometrial carcinoma. Basic.LncRNA is a transcriptional transcript with a length of more than 200nt for proteins that can not be encoded. A large number of studies have found that it plays an important role in human life activities and the development of various diseases..CARLo-5 is a recently discovered lncRNA of 2628 nucleotides, located near the enhancer of the proto oncogene myc. It is reported that CARLo-5 is in the colon. Cancer, gastric cancer and other cancer tissues are up-regulated and play an important role in the development and progression of cancer. The diagnosis, evaluation and treatment of various cancers are of potential value. However, the expression and mechanism of CARLo-5 in endometrial carcinoma have not yet been reported. This study uses real-time fluorescent quantitative PCR (Real-time PCR). The expression of CARLo-5 gene in normal endometrium and endometrial carcinoma tissues was detected. The expression of cell cycle related proteins CDK2 and CDKN1A were detected by Western blot (Western-blot). The correlation between the expression level of CARLo-5 and the clinicopathological features and the total survival time of endometrial cancer patients was analyzed. The correlation between the expression level of CARLo-5 gene and the expression level of cell cycle related protein CDK2 and CDKN1A was analyzed. After the expression of CARLo-5 in low endometrial carcinoma cells in vitro, MTS, Transwell migration, Matrigel invasion test, flat clone formation experiment and flow cytometry were used to detect the biological line of endometrial cancer by knocking low CARLo-5 to endometrial carcinoma. In order to explore the mechanism of CARLo-5 in endometrial carcinoma, this study examined the effect of Western-blot on the expression of CDK2, CDKN1A, matrix metalloproteinase and MMP9 in endometrial carcinoma cells by knocking low CARLo-5. Through RNAi lentivirus technology, we constructed a stable knockout low CARLo-5 endometrial carcinoma cell line. The effect of knockout CARLo-5 on the proliferation of endometrial carcinoma cells in nude mice was tested. This study explored the role and mechanism of CARLo-5 in endometrial carcinoma in order to elucidate the molecular mechanism of endometrial cancer and the genetic diagnosis and treatment of endometrial carcinoma. The expression of CARLo-5 in endometrial carcinoma and its clinical significance: To investigate the expression of lncRNA CARLo-5 in endometrial carcinoma and to explore the relationship between the expression of CARLo-5 and the clinicopathological features and the total survival period of endometrial carcinoma. Expression level of cyclin related protein CDK2 and CDKN1A and their correlation with CARLo-5 expression. Methods: 1 the expression level of CARLo-5 in 108 endometrial carcinoma tissues and 66 normal endometrium tissues was detected by Real-time PCR, and Western-blot was used to detect the cell cycle phase in endometrial carcinoma and normal endometrium by Western-blot Expression of protein CDK2 and CDKN1A.3 analysis of the correlation between the expression level of CARLo-5 and the clinicopathological features and the total survival period of endometrial carcinoma.4 analysis of the correlation between the expression of CARLo-5 in endometrial carcinoma and the expression of cell cycle related protein CDK2 and CDKN1A. Fruit: 1 lncRNA CARLo-5 in endometrial carcinoma tissue The expression level of Real-time PCR in endometrium and normal endometrium showed that the expression level of CARLo-5 in endometrial carcinoma was significantly higher than that of normal endometrium (P0.001), the expression level of.2 CARLo-5 and the clinicopathological features of endometrial carcinoma patients, the expression level of CARLo-5 in endometrial carcinoma tissue The clinical stage of endometrial carcinoma is closely related to lymph node metastasis (P0.05). There is no significant correlation with age, pathological type, pathological grade, muscular layer invasion depth and vascular infiltration (P0.05), the correlation between the expression level of.3 CARLo-5 and the total survival period of endometrial cancer patients with endometrial carcinoma, the average of CARLo-5 high expression group of endometrium cancer patients The survival time (43 + 14.1 months) was significantly shorter than that of the CARLo-5 low expression group (52.5 + 9.1 months). The expression level of CDK2 and CDKN1A protein in endometrial carcinoma tissue of (P0.05).4 showed that the expression level of CDK2 protein in endometrial carcinoma (9.32 + 0.69) was significantly higher than that of normal endometrium (1 + 0.54), and CDKN1A The expression level of protein in endometrial carcinoma (0.71 + 0.14) was significantly lower than that of normal endometrium (1 + 0.11). The expression level of.5 CARLo-5 and the expression level of CDK2 and CDKN1A protein were correlated with the expression level of CDK2 protein in endometrial carcinoma and the expression of CARLo-5 was positively correlated (r=0.30, P=0.043).CDKN1A. The expression of protein was negatively correlated with the expression of CARLo-5 (r=-0.28, P=0.046). Conclusion: the expression of 1 CARLo-5 in endometrial carcinoma is significantly higher than that of normal endometrium, suggesting that CARLo-5 may play a role in the development of endometrial carcinoma, which may promote the clinical stage of.2 CARLo-5 high and endometrial cancer patients. Metastasis is associated with poor prognosis, suggesting that CARLo-5 may be a molecular marker for predicting the prognosis of endometrial carcinoma. The expression level of.3 CDK2 protein in endometrial carcinoma is significantly higher than that of normal endometrium, and the expression of CDKN1A protein is significantly reduced, and the expression of CDKN1A and CDK2 protein is significantly related to the expression level of CARLo-5. These results suggest that CARLo-5 may play a role in promoting cancer by regulating the expression of CDK2 and CDKN1A protein in endometrial carcinoma. Second the effect of part CARLo-5 on the biological behavior of endometrial carcinoma cells and its mechanism: To investigate the expression of CARLo-5 in endometrial carcinoma cells to explore the intrauterine The effects of membrane cancer cell biological behavior and its possible mechanisms. Methods: 1 the effects of MTS on the proliferation of endometrial carcinoma cells in vitro were detected by MTS method. Transwell migration, scratch healing experiment and Matrigel invasion test were used to detect the effect of CARLo-5 on the migration and invasion ability of endometrial carcinoma cells in vitro and.3 mining. The effect of knockout low CARLo-5 on the clone formation ability of endometrial cancer cells using a flat cloned clone.4 the effect of.4 on the proliferation cycle of endometrial carcinoma cells by flow cytometry;.5 and Real-time PCR and Western-blot were used to detect the CDK2 and CDKN1A genes and protein tables in endometrial cancer cells with Real-time PCR and Western-blot Effect of.6 on the expression of MMP2 and MMP9 protein in endometrial carcinoma cells by Western-blot. Results: 1 the effect of knockout low CARLo-5 on the proliferation activity of endometrial carcinoma cells in vitro MTS assay results showed that the proliferation activity of HEC-1-B and KLE in endometrial carcinoma cells after knocking low CARLo-5 was obvious. The effect of reduced (P0.05).2 knockout CARLo-5 on the migration and invasion ability of endometrial carcinoma cells in vitro, Transwell migration and scratch healing experiment showed that the migration ability of HEC-1-B and KLE in endometrial carcinoma cells decreased significantly after the knock at low CARLo-5 (P0.05); Matrigel invasion experiment results showed that endometrial cancer cells were H after low CARLo-5 The invasiveness of EC-1-B and KLE in vitro significantly decreased (P0.05).3 knockdown CARLo-5 effect on the clone formation ability of endometrial carcinoma cells. Experimental results showed that the clone formation ability of HEC-1-B and KLE in endometrial carcinoma cells decreased significantly after low CARLo-5 (P0.05).4 knockdown CARLo-5 on the proliferation cycle of endometrial carcinoma cells The results of flow cytometry showed that the number of G0/G1 cells in the endometrial carcinoma HEC-1-B and KLE cells increased significantly after the knockout of CARLo-5, and the number of cells in the S phase decreased significantly (P0.05) the effect of.5 knocking low CARLo-5 on the gene and protein expression level of CDK2 and CDKN1A in endometrial carcinoma cells The expression level of CDK2 m RNA and protein in endometrium cancer cells decreased significantly (P0.05), while the expression level of CDKN1A m RNA and protein increased significantly (P0.05).6 knock low CARLo-5 on endometrial carcinoma cells MMP2 and MMP9 protein expression level P0.05. Conclusion: 1 knockout low CARLo-5 can inhibit the proliferation, migration and invasion ability of endometrial carcinoma cells in vitro, suggesting that CARLo-5 expression can promote the proliferation, invasion and metastasis of endometrial cancer cells. CARLo-5 is expected to be a new biological target for the treatment of endometrial cancer,.2 knockdown CARLo-5 to make endometrial cancer cells Blocking the expression of CDK2 m RNA and protein levels and promoting the expression of CDKN1A m RNA and protein levels, suggesting that CARLo-5 may inhibit the G0/G1 phase block by regulating the expression of CDK2 and CDKN1A, and then promote the proliferation of endometrial cancer cells to inhibit the proliferation of endometrial carcinoma cells and inhibit the G0/G1 and the protein of endometrial cancer cells. Expression, suggesting that CARLo-5 may promote the metastasis of endometrial carcinoma by up regulation of the expression of MMP2 and MMP9 protein. Third the effect of low CARLo-5 on the growth of nude mice with endometrial cancer cells and its mechanism: to detect the effect of low CARLo-5 on the tumorigenicity of endometrial cancer cells in nude mice The mechanism of action in the development of endometrial carcinoma in mice in vivo. Methods: 1 the effect of RNAi Lentivirus on endometrial cancer cell line that stably knockdown CARLo-5 was constructed in.2 nude mice, the effects of low CARLo-5 on the tumorigenicity of endometrial carcinoma cells were detected by.3 immuno histochemical test in nude mice The expression level of CDK2 and CDKN1A protein in the transplanted tumor tissue. Results: 1 the effect of low CARLo-5 on the endometrial cancer cells in the body sh RNA-1 group, the average volume of the transplanted tumor in the SH RNA-2 group and the control group was (157.66 + 37.93 mm3), (102.81 + 58.33mm3), (499.74 + 98.61 mm3), and the growth rate of nude mice in the low CARLo-5 group was the growth rate and the growth rate of the nude mice. The effect of.2 knock low CARLo-5 on the expression level of CDK2 and CDKN1A protein in transplanted tumor tissues of nude mice was significantly lower than that in the lower group (P0.05). The immunohistochemical results showed that the protein expression level of CDK2 in the transplanted tumor tissues of the low CARLo-5 group was significantly lower than that in the lower group (P0.05), while the level of the CDKN1A protein expression was not knockout. The lower group was significantly increased (P0.05). Conclusion: the knock down CARLo-5 obviously inhibited the proliferation of endometrial carcinoma cells in vivo. The mechanism may be that CARLo-5 inhibits CDKN1A and promotes the expression of CDK2 protein, thus promoting the proliferation of endometrial cancer cells in vivo. CARLo-5 may be a new target for the treatment of endometrium carcinoma.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R737.33

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1 趙喜娃;CARLo-5在子宮內(nèi)膜癌中的作用及機(jī)制研究[D];河北醫(yī)科大學(xué);2017年

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