蛋白激酶CK2α在卵巢癌中的表达及作用机制探讨
发布时间:2018-06-16 18:02
本文选题:蛋白激酶CK2α + β-Catenin ; 参考:《辽宁医学院》2014年硕士论文
【摘要】:目的 通过研究蛋白激酶CK2α在卵巢癌组织和OVCAR3细胞株中的表达情况,分析CK2α与β-Catenin、Survivin在卵巢癌中的相互作用关系,进一步探讨蛋白激酶CK2α在卵巢癌发生发展中的作用机制。 方法 1、采用免疫组化SP法检测CK2α、β-Catenin、Survivin在8例卵巢单纯性囊肿、8例卵巢良性囊腺瘤、20例卵巢浆液性囊腺癌,9例卵巢粘液性囊腺癌组织标本中的表达情况,并分析CK2α、β-Catenin、Survivin在卵巢癌中的相关性。 2、体外培养人卵巢癌OVCAR3细胞株,实验分为空白对照组(未处理组)、DMSO溶剂组、TBB组(TBB终浓度分别为12.5μmol/L、25μmol/L、50μmol/L、75μmol/L、100μmol/L)。药物作用24h后,MTT比色法测定细胞生长抑制率。 3、应用RT-PCR和Western blotting技术分别检测蛋白激酶CK2α、β-Catenin、Survivin mRNA和蛋白在各组人卵巢癌OVCAR3细胞中的表达,分析各组CK2α和β-Catenin、Survivin基因表达的变化及它们之间的相互关系。 结果 1、与卵巢单纯性囊肿和良性卵巢肿瘤组织相比,CK2α、β-Catenin、Survivin在卵巢癌中的蛋白表达水平明显提高P0.05㖞;CK2α与β-Catenin、Survivin在卵巢癌中表达呈正相关(R=0.438、0.479,P0.05);CK2α、β-Catenin、Survivin在低 中分化组的蛋白表达水平阳性率明显高于高分化组(P0.05);CK2α、β-Catenin、Survivin在Ⅱ Ⅲ期的蛋白表达水平阳性率与Ⅰ期相比明显增加,差异有统计学意义(P0.05)。 2、与DMSO溶剂组相比,以不同浓度TBB组处理的OVCAR3细胞生长抑制率随TBB浓度的增加而明显增加,呈剂量依赖性。 3、RT-PCR结果表明,随着TBB浓度的增加,OVCAR3细胞株CK2α的mRNA表达水平降低,但β-Catenin、Survivin的mRNA表达水平并没有明显变化。 4、Western blotting结果表明,与对照组相比,加入TBB后,CK2α的蛋白表达受到高效特异的抑制,β-Catenin、Survivin蛋白表达水平明显降低。 结论 1、蛋白激酶CK2α、β-Catenin、Survivin高表达可能与卵巢癌的发生和发展有关,并且CK2α与β-Catenin、Survivin在卵巢癌中的表达呈正相关 2、TBB对人卵巢癌OVCAR3细胞株细胞生长增殖具有显著抑制作用,其机制可能与降低CK2α基因表达,进而下调细胞增殖、凋亡相关基因β-Catenin、Survivin蛋白表达水平与活性有关。
[Abstract]:Objective to study the expression of protein kinase CK2 伪 in ovarian carcinoma and OVCAR3 cell line, and to analyze the interaction between CK2 伪 and 尾 -Catenin survivin in ovarian cancer, and to explore the mechanism of protein kinase CK2 伪 in the development of ovarian cancer. Methods 1. Immunohistochemical SP method was used to detect the expression of CK2 伪, 尾 -Catenin survivin in 8 cases of ovarian simple cysts and 8 cases of benign ovarian cystadenoma, 20 cases of ovarian serous cystadenocarcinoma and 9 cases of ovarian mucinous cystadenocarcinoma. The correlation of CK2 伪, 尾 -Catenin survivin in ovarian cancer was analyzed. 2Human ovarian cancer OVCAR3 cell line was cultured in vitro and divided into blank control group (the final concentration of TBB in untreated group was 12.5 渭 mol / L ~ 25 渭 mol / L ~ 50 渭 mol / L ~ (75) 渭 mol 路L ~ (-1) ~ 100 渭 mol 路L ~ (-1). The cell growth inhibition rate was measured by MTT colorimetry 24 hours after treatment. The expression of protein kinase CK2 伪, 尾 -Catenin survivin mRNA and protein in human ovarian cancer OVCAR3 cells was detected by RT-PCR and Western blotting. The expression changes of CK2 伪 and 尾 -Catenin survivin gene in each group and their relationship were analyzed. Results (1) compared with simple ovarian cyst and benign ovarian tumor, the expression of CK2 伪, 尾 -Catenin survivin in ovarian carcinoma was significantly higher than that in ovarian carcinoma. The expression of P0.05a and 尾 -Catenin survivin in ovarian carcinoma was positively correlated with the expression of 尾 -Catenin survivin in ovarian carcinoma. The positive rate of CK2 伪, 尾 -Catenin survivin protein expression in poorly differentiated group was significantly higher than that in well-differentiated group (P 0.05). The positive rate of 尾 -Catenin survivin protein expression in stage 鈪,
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