【摘要】：目的：子宮內(nèi)膜癌是女性生殖系統(tǒng)最常見的惡性腫瘤，子宮內(nèi)膜癌的發(fā)生、發(fā)展與很多因素有關(guān)，如長期雌激素刺激同時無孕激素拮抗等機(jī)制。同時，他莫昔芬（tamoxifen TAM）作用后，子宮內(nèi)膜癌的發(fā)生率明顯升高。但是，目前對于TAM引起子宮內(nèi)膜癌的發(fā)病機(jī)制仍不清楚。關(guān)于染色體重組復(fù)合物的研究日益增多，具有抑制DNA合成的作用，故其突變低表達(dá)能夠促進(jìn)細(xì)胞無限分裂，進(jìn)而發(fā)展成癌。ARID1A（AT-richinteracting domain containing protein，又稱腦蛋白120、SMARCF1、p270等）是染色質(zhì)重組復(fù)合物之一，同時作為抑癌基因，在多種腫瘤組織中存在突變。目前對于ARID1A的結(jié)構(gòu)研究較多，對其在子宮內(nèi)膜癌中的表達(dá)及生物學(xué)功能的研究較少。本研究通過檢測ARID1A、ERα、ERβ、PR、P53在子宮內(nèi)膜癌細(xì)胞（高分化Ishikawa、中分化HEC-1A及低分化KLE）中的基因和蛋白的表達(dá)水平，并進(jìn)一步分析TAM及雌激素（estrogen E2）處理后子宮內(nèi)膜癌細(xì)胞中ARID1A、ERα、ERβ、PR、P53基因和蛋白的表達(dá)水平變化，探討ARID1A在子宮內(nèi)膜癌的發(fā)生、發(fā)展中的可能作用機(jī)制。 方法： 1應(yīng)用實時定量qRT-PCR法檢測正常子宮內(nèi)膜組織、子宮內(nèi)膜癌細(xì)胞株（Ishikawa、HEC-1A、KLE）中ARID1A、ERα、ERβ、PR、P53mRNA的表達(dá)水平。 2應(yīng)用實時定量qRT-PCR法檢測TAM及E2依據(jù)不同濃度及不同時間處理細(xì)胞后ARID1A、ERα、ERβ、PR、P53mRNA的表達(dá)水平。 3應(yīng)用Western-blot法檢測正常子宮內(nèi)膜組織、子宮內(nèi)膜癌細(xì)胞株（Ishikawa、HEC-1A、KLE）中BAF250a、ERα、ERβ、PR、P53蛋白的表達(dá)水平。 4應(yīng)用Western-blot法檢測TAM及E2依據(jù)不同濃度及不同時間處理細(xì)胞后BAF250a、ERα、ERβ、PR、P53蛋白的表達(dá)水平。 5應(yīng)用流式細(xì)胞學(xué)測定TAM及E2依據(jù)不同濃度及不同時間處理細(xì)胞后細(xì)胞周期變化情況。 結(jié)果： 1ARID1A、ERα、ERβ mRNA及蛋白在三種子宮內(nèi)膜癌細(xì)胞中的表達(dá)均顯著低于其在正常子宮內(nèi)膜組織中的表達(dá)，經(jīng)過TAM或E2處理后，ARID1A、ERα、ERβ mRNA及蛋白表達(dá)量隨著藥物濃度的增加而降低，隨著藥物處理時間的延長而降低(P 0.05)，而PR、P53mRNA及蛋白在三種子宮內(nèi)膜癌細(xì)胞中的表達(dá)低于其在正常子宮內(nèi)膜組織中的表達(dá)，經(jīng)過TAM或E2處理后，PR、P53mRNA及蛋白表達(dá)量隨著藥物濃度的增加而增加，隨著藥物處理時間的延長而增加，但均低于其在正常子宮內(nèi)膜組織中的表達(dá)(P 0.05)，呈時間-劑量依賴性。 2經(jīng)過不同濃度TAM或E2在不同時間處理三種子宮內(nèi)膜癌細(xì)胞（Ishikawa、HEC-1A、KLE）后，三種細(xì)胞均表現(xiàn)為G0/G1期縮短，S期延長，且隨著藥物濃度的增高及處理時間的延長，G0/G1期逐漸縮短，S期逐漸延長（P0.05）。 結(jié)論： 1TAM或E2處理子宮內(nèi)膜癌細(xì)胞后ARID1A、ERα、ERβmRNA及蛋白的表達(dá)明顯低于其在未經(jīng)藥物處理子宮內(nèi)膜癌中的表達(dá)，而PR、P53mRNA及蛋白的表達(dá)明顯高于其在未經(jīng)藥物處理子宮內(nèi)膜癌中的表達(dá)。 2TAM與E2可能通過下調(diào)ARID1A的表達(dá)而引起子宮內(nèi)膜癌的發(fā)生，，而且ARID1A的下調(diào)可能與ER、P53存在一定相互作用關(guān)系。 3經(jīng)TAM與E2處理后子宮內(nèi)膜癌細(xì)胞的細(xì)胞周期均表現(xiàn)為G0/G1期縮短，S期延長。
[Abstract]:Objective: Endometrial carcinoma is the most common malignant tumor of female reproductive system. The occurrence and development of endometrial carcinoma are related to many factors, such as long-term estrogen stimulation and no such mechanism as progestogen antagonism. At the same time, tamoxifen (tamoxifen) was used to increase the incidence of endometrial carcinoma. However, the pathogenesis of endometrial cancer is still unclear for TAM. The research on the chromosomal recombination complex has been increasing, and has the effect of inhibiting the DNA synthesis, so that the mutant low expression can promote the infinite division of the cell, and further develop the cancer. ARID1A (AT-ricinting domain containment protein, also known as brain protein 120, SMARCF1, p270, etc.) is one of the recombinant complexes of chromatin, and as a tumor suppressor gene, there are mutations in various tumor tissues. At present, there are many studies on the structure of ARID1A, and the study of its expression and biological function in endometrial carcinoma is less. The expression of ARID1A, ER, ER, PR, P53 in endometrial cancer cells (HEC-1A and low-differentiated KLE) and the expression levels of genes and proteins in endometrial cancer cells (HEC-1A and low-differentiated KLE) were detected and the ARID, ER, ER and ER levels in the endometrial cancer cells were further analyzed. The expression level of PR, P53 gene and protein was changed, and the possible mechanism of ARID1A in the development and development of endometrial carcinoma was discussed. square Method:1 A real-time quantitative qRT-PCR was used to detect the expression of ARID1A, ER, ER, PR, P53mRNA in normal endometrium and endometrial carcinoma (Ishikawa, HEC-1A, KLE). The levels of AID1A, ER, ER, PR, P53mRNA were detected by real-time quantitative qRT-PCR. The level of expression of BAF250a, ER, ER, PR, P53 in normal endometrial tissue, endometrial carcinoma cell (Ishikawa, HEC-1A, KLE) was detected by Western-blot. The expression level of the protein was detected by Western-blot, and then the cells were treated with different concentrations and different time. The BAF250a, ER antigen, ER antigen, PR, P were treated with different concentrations and time. The expression level of 53 protein was measured by flow cytometry. After the cell Results: The expression of ARID1A, ER, ER, ER-mRNA and protein in three endometrial cancer cells was significantly lower than that in normal endometrial tissues. The expression of PR, P53mRNA and protein in the three endometrial cancer cells decreased with the increase of the drug concentration (P 0.05). The expression of PR, P53mRNA and protein in the three endometrial cancer cells was lower than that in the normal endometrial tissues. The expression of PR, P53mRNA and protein after treatment with TAM or E2 The amount of expression increases with the increase of the drug concentration, which increases with the increase of the drug treatment time, but is lower than the expression of the drug in the normal endometrium (P 0.05) Time-dose-dependent. After treatment of three endometrial cancer cells (Ishikawa, HEC-1A, KLE) at different times through different concentrations of TAM or E2, the three cells showed a decrease in G0/ G1 phase and an extension of S phase, and the G0/ G1 phase was decreased with the increase of drug concentration and the time of treatment. gradual shortening Conclusion: The expression of ARID1A, ER, ER, ER-mRNA and protein in the endometrial carcinoma cells after treatment with 1TAM or E2 was significantly lower than that of the non-drug-treated endometrial carcinoma, while the expression of PR, P53mRNA and protein was significantly lower. significantly higher than that in endometrial cancer without drug treatment. TAM and E2 may cause endometrial cancer by down-regulation of the expression of ARID1A, and ARID1 A's down-regulation may have a certain interaction with ER and P53. After treatment with TAM and E2, the endometrium
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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