本文選題：卵巢癌 + 組蛋白去甲基化轉(zhuǎn)移酶KDMB��； 參考：《現(xiàn)代婦產(chǎn)科進(jìn)展》2017年03期

【摘要】：目的:檢測組蛋白去甲基化轉(zhuǎn)移酶KDM5B(JARID1B/PLU-1)在正常卵巢、卵巢癌組織及細(xì)胞株中的表達(dá),以及其對卵巢癌細(xì)胞株耐藥能力的影響。方法:RT-q PCR法檢測KDM5B在36例卵巢癌、15例正常卵巢、6例術(shù)后化療耐藥復(fù)發(fā)患者組織及卵巢上皮細(xì)胞株IOSE和5株卵巢癌細(xì)胞中的表達(dá)情況;構(gòu)建p MSCVpuro-KDM5B過表達(dá)質(zhì)粒和p GIPZ-sh1、p GIPZ-sh2干擾質(zhì)粒,并篩選得到穩(wěn)定過表達(dá)的SKOV3和穩(wěn)定干擾的Caov3細(xì)胞株,RT-q PCR和Western blot法鑒定調(diào)控效果;用濃度梯度遞增的順鉑處理SKOV3過表達(dá)和Caov3干擾細(xì)胞株,CCK-8法檢測順鉑半抑制濃度(IC50)變化。結(jié)果:RTq PCR結(jié)果示,正常卵巢組織中KDM5B相對表達(dá)量(1.950±0.3003)低于卵巢癌組織(4.924±0.2989),差異有統(tǒng)計學(xué)意義(t=5.909,P0.0001)。KDM5B表達(dá)量與卵巢癌患者的FIGO分期、病理分級、轉(zhuǎn)移和耐藥發(fā)生明顯相關(guān),與患者年齡、CA125水平不相關(guān)。6例復(fù)發(fā)患者中,5例KDM5B表達(dá)明顯升高(P0.001)。KDM5B的mRNA和蛋白水平在卵巢上皮性細(xì)胞株IOSE中最低,在卵巢癌細(xì)胞株SKOV3、HO-8910、ES-2、A2780、Caov3中表達(dá)逐漸增加。RT-q PCR結(jié)果示,SKOV3過表達(dá)組的KDM5B表達(dá)量為對照組的27.4倍,Caov3干擾sh1和sh2組分別為對照組的0.38和0.41倍,Western blot也顯示過表達(dá)和干擾有效。用濃度梯度遞增的順鉑處理Caov3細(xì)胞株,KDM5B表達(dá)量隨順鉑濃度增加而逐漸升高;檢測SKOV3細(xì)胞株KDM5B過表達(dá)組IC50為3.666(95%CI為3.067~4.382)μg/ml高于對照組[1.676(95%CI為1.553~1.810)μg/ml],Caov3細(xì)胞株干擾sh1組和sh2組分別為3.359(95%CI為2.875~3.925)μg/ml、2.881(95%CI為2.49~3.324)μg/ml,均低于對照組[6.972(95%CI為6.182~7.864)]。結(jié)論:組蛋白去甲基化轉(zhuǎn)移酶KDM5B在卵巢癌中表達(dá)升高,并且具有促進(jìn)卵巢癌細(xì)胞株耐藥的作用。
[Abstract]:Aim: to detect the expression of histone demethyltransferase (KDM5BnJARID1B / PLU-1) in normal ovary, ovarian cancer tissues and cell lines, and its effect on the drug resistance of ovarian cancer cell lines. Methods the expression of KDM5B in 36 ovarian cancer tissues, ovarian epithelial cell lines IOSE and 5 ovarian cancer cell lines were detected by RT-PCR, and the overexpression plasmids of MSCP Vpuro-KDM5B and pGIPZ-sh1pGIPZ-sh2 interference plasmids were constructed. Stable overexpression of SKOV3 and stable interference of Caov3 cell line RT-q PCR and Western blot assay were used to identify the regulatory effect, and Cisplatin was used to treat SKOV3 overexpression and Caov3 interference cell line CCK-8 to detect the change of Cisplatin semi-inhibitory concentration (IC50). Results the relative expression of KDM5B in normal ovarian tissues (1.950 鹵0.3003) was significantly lower than that in ovarian cancer (4.924 鹵0.2989). There was a significant correlation between the expression of KDM5B and Figo stage, pathological grade, metastasis and drug resistance in patients with ovarian cancer. The expression of KDM5B in 5 patients with recurrent ovarian epithelial cell line IOSE was the lowest in ovarian epithelial cell line IOSE, and the mRNA and protein levels of KDM5B were significantly higher in 5 patients than those in ovarian epithelial cell line IOSE. The expression of KDM5B in ovarian cancer cell line SKOV3HO-8910FES-2A2780 was gradually increased. RT-q PCR results showed that the overexpression of KDM5B in SKOV3 overexpression group was 27.4 times as much as that in control group. Caov3 interfered with sh1 and sh2 group was 0.38 and 0.41 times of the control group respectively. The overexpression and interference of KDM5B in SKOV3 overexpression group were also shown to be effective. The expression of KDM5B in Caov3 cell line increased with the increase of cisplatin concentration. The IC50 of the overexpression group of SKOV3 cell line KDM5B was 3.666N95 CI 3.0674.382) 渭 g/ml, which was higher than that of the control group [1.676995 CI was 1.5531.810 渭 g/ml] Caov3 cell line interfered with sh1 group and sh2 group with 3.35995CI of 2.875c3.925) 渭 g ml / ml, which was lower than that of the control group (6.97295CI was 6.1827.864). Conclusion: the expression of histone demethyltransferase KDM5B is increased in ovarian cancer and can promote the drug resistance of ovarian cancer cell line.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院婦產(chǎn)科;
【基金】：國家自然科學(xué)基金項(xiàng)目(No:81572560)
【分類號】：R737.31
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本文編號：2021419