KDM5B基因在卵巢癌中的表达及其对卵巢癌耐药能力的影响

发布时间：2018-06-15 08:35

  本文选题：卵巢癌 + 组蛋白去甲基化转移酶KDMB　； 参考：《现代妇产科进展》2017年03期

【摘要】：目的:检测组蛋白去甲基化转移酶KDM5B(JARID1B/PLU-1)在正常卵巢、卵巢癌组织及细胞株中的表达,以及其对卵巢癌细胞株耐药能力的影响。方法:RT-q PCR法检测KDM5B在36例卵巢癌、15例正常卵巢、6例术后化疗耐药复发患者组织及卵巢上皮细胞株IOSE和5株卵巢癌细胞中的表达情况;构建p MSCVpuro-KDM5B过表达质粒和p GIPZ-sh1、p GIPZ-sh2干扰质粒,并筛选得到稳定过表达的SKOV3和稳定干扰的Caov3细胞株,RT-q PCR和Western blot法鉴定调控效果;用浓度梯度递增的顺铂处理SKOV3过表达和Caov3干扰细胞株,CCK-8法检测顺铂半抑制浓度(IC50)变化。结果:RTq PCR结果示,正常卵巢组织中KDM5B相对表达量(1.950±0.3003)低于卵巢癌组织(4.924±0.2989),差异有统计学意义(t=5.909,P0.0001)。KDM5B表达量与卵巢癌患者的FIGO分期、病理分级、转移和耐药发生明显相关,与患者年龄、CA125水平不相关。6例复发患者中,5例KDM5B表达明显升高(P0.001)。KDM5B的mRNA和蛋白水平在卵巢上皮性细胞株IOSE中最低,在卵巢癌细胞株SKOV3、HO-8910、ES-2、A2780、Caov3中表达逐渐增加。RT-q PCR结果示,SKOV3过表达组的KDM5B表达量为对照组的27.4倍,Caov3干扰sh1和sh2组分别为对照组的0.38和0.41倍,Western blot也显示过表达和干扰有效。用浓度梯度递增的顺铂处理Caov3细胞株,KDM5B表达量随顺铂浓度增加而逐渐升高;检测SKOV3细胞株KDM5B过表达组IC50为3.666(95%CI为3.067~4.382)μg/ml高于对照组[1.676(95%CI为1.553~1.810)μg/ml],Caov3细胞株干扰sh1组和sh2组分别为3.359(95%CI为2.875~3.925)μg/ml、2.881(95%CI为2.49~3.324)μg/ml,均低于对照组[6.972(95%CI为6.182~7.864)]。结论:组蛋白去甲基化转移酶KDM5B在卵巢癌中表达升高,并且具有促进卵巢癌细胞株耐药的作用。
[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 卤0.3003) was significantly lower than that in ovarian cancer (4.924 卤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.
【作者单位】： 上海交通大学医学院附属仁济医院妇产科;
【基金】：国家自然科学基金项目(No:81572560)
【分类号】：R737.31
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本文编号：2021419