P65、P50及IκB-α蛋白表达水平与人卵巢癌细胞化疗耐药性关系研究

Expressions of P65, P50 and IκB-α Proteins and its Relationship with drug resistance in human ovarian cell lines

目的探讨P65、P50及IκB-α蛋白表达水平与人卵巢癌细胞化疗耐药性关系。方法培养10株卵巢癌细胞系,当各株卵巢癌细胞生长到对数生长期时,取细胞进行细胞质、细胞核P65、P50及IκB-α蛋白质提取。Western blot检测P65、P50及IκB-α蛋白质表达情况,采用MTT法对10株卵巢癌细胞系进行了IC50值检测、分析P65、P50蛋白与药物敏感性的关系。结果 P65、P50及IκB-α蛋白主要在细胞质中表达,P65、P50及IκB-α蛋白在细胞质中表达阳性率分别为80.00%、60.00%、70.00%;P65、P50蛋白也在细胞核中表达,P65、P50蛋白在细胞核中表达阳性率分别为20.00%、50.00%,IκB-α蛋白未见表达于细胞核中。OV-MZ-5细胞系具有最高的IC50值,为20.34μg/ml;OV-MZ-12b细胞系具有最低的IC50值,为4.23μg/mL。各组卵巢癌细胞系IC50值未显示出明显规律。P65胞核阳性者IC50平均值(17.23μg/mL)明显高于胞核阴性者IC50平均值(8.45μg/mL),且高出后者1倍以上;而P50胞核阳性IC50平均值(9.83μg/mL)与胞核阴性者IC50平均值(10.22μg/mL)相近。结论 P65和P50蛋白在卵巢癌细胞的发生、发展中具有重要作用,并且P65的表达水平与卵巢癌细胞的化疗耐药性息息相关,有望成为判断预后的生物标志物。

Objective To investigate the expressions of P65, P50 and IκB-α Proteins and its Relationship with drug resistance in human ovarian cell lines. Methods All 10 ovarian cancer cell lines were cultured, and then P65, P50 and IκB-αproteins were extracted separately from cytoplasm or nuclear of 10 ovarian cancer cell lines. The expressions of P65, P50 and IκB-α proteins were detected by using western blotting. IC50 of 10 ovarian cancer cell lines were detected by using MTT drug resistance testing. The relationship between the expressions of P65, P50 and IκB-αproteins in ovarian cancer cell lines and chemotherapy resistance of ovarian cell was observed. Results The expressions of P65, P50 and IκB-α proteins were distributed in cytoplasm, and the expression of positive rate were 80.00%, 60.00% and 70.00%, respectively. The expression of positive rate of P65 and P50 in nuclear were20.00% and 50.00%, respectively. There was no IκB-α expression in nuclear. In OV-MZ-5 ovarian cancer cell line, the expression of P65 in nuclear was positive and it's value of IC50(17.23 μg/mL) was the highest, and it's value of IC50 was more than twice as much as the value of IC50 in ovarian cancer cell line which the expression of P65 in nuclear was negative. In OV-MZ-5 ovarian cancer cell line, the expression of P65 in nuclear was positive and it's value of IC50(17.23 μg/mL) was the highest. In OV-MZ-12 b ovarian cancer cell line, the expression of P65 in nuclear was positive and it's value of IC50(4.23μg/mL) was the lowest. The value of IC50(17.23 μg/mL) in P65 expression positive cells was more than twice as much as the value of IC50(8.45 μg/ml) in P65 expression negative cells. The value of IC50 of P50 had no definite trend in expression positive cells(9.83 μg/mL) and negative cells(10.22 μg/mL). Conclusion P65, P50 play an important role in the progress of ovarian cancer cells. There existed a significant correlation between the expressions of P65, P50 and the chemotherapy resistance of ovarian cancer cells. and these could be expected to be the new markers in the observation of prognosis.