CARD9蛋白与乳腺癌患者特征及癌细胞增殖、凋亡、侵袭的影响

Effect of CARD9 Protein on the Characteristics of Breast Cancer Patients and the Proliferation,Apoptosis and Invasion of the Cancer Cells

目的探讨胱天蛋白酶募集域蛋白9(caspase recruitment domain containing protein 9,CARD9)与乳腺癌患者临床特征关系及对乳腺癌MDA-MB-231细胞增殖、凋亡、侵袭的影响。方法选取2018年1月至2019年9月咸宁市中心医院保存的乳腺癌标本90例,同时选取癌旁组织作为对照,采用Western印迹检测CARD9蛋白表达,组间差异比较采用t检验;选取人类乳腺癌细胞株MDA-MB-231细胞,分为对照组、空白转染组和阳性转染组,其中空白转染组转染空白质粒,阳性转染组转染CARD9表达质粒,采用CCK8试验检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell试验检测细胞侵袭,组间差异比较采用F检验。结果乳腺癌组织CARD9蛋白相对表达量为(1.03±0.22),明显高于癌旁组织(P<0.05);肿瘤直径>5 cm、TNM分期Ⅲ期、ER表达阳性组织CARD9蛋白相对表达量分别为(1.12±0.20)、(1.27±0.21)和(1.12±0.19),明显高于肿瘤直径≤5 cm、TNM分期Ⅰ~Ⅱ期、ER表达阴性组织(P<0.05);阳性转染组细胞培养24、48和72 h时A值分别为(0.487±0.102)、(0.702±0.107)和(0.972±0.106),明显高于对照组和空白转染组(P<0.05);阳性转染组、空白转染组和对照组细胞凋亡率比较差异无统计学意义(P>0.05);阳性转染组细胞侵袭数为(101.2±11.4)个,明显多于空白转染组和对照组(P<0.05)。结论乳腺癌组织CARD9蛋白表达上调,与肿瘤直径、TNM分期及ER表达有一定关系;CARD9表达对乳腺癌MDA-MB-231细胞增殖及侵袭能力有一定影响,对细胞凋亡无明显影响。

Objective To investigate the relationship between Caspase recruitment domain containing protein 9(CARD9)and the clinical characteristics of breast cancer patients and the effects of CARD9 on the proliferation,apoptosis and invasion of MDA-MB-231 cells.Methods 90 breast cancer specimens preserved in our hospital from January 2018 to September 2019 were examined,with the adjacent tissues serving as controls.The CARD9 protein expression was detected by Western blotting and t test was employed to compare the difference between groups.Human breast cancer cell line MDA-MB-231 were divided into a control group,a blank transfection group and a positive transfection group.The blank transfection group was transfected with blank plasmids and the positive transfection group was transfected with CARD9-expressing plasmids.The cell proliferation was detected by CCK8 test;cell apoptosis was detected by flow cytometry;cell invasion was detected by Transwell test;F test was used to compare the differences between groups.Results The relative expression level of CARD9 protein in breast cancer tissues was(1.03±0.22),which was significantly higher than that in adjacent tissues(P<0.05).The relative expression of CARD9 protein in tissues with tumor diameter>5 cm,TNM stage El,ER positive expression were(1.12±0.20),(1.27±0.21)and(1.12±0.19),which were significantly higher than in tissues with the tumor diameter<5 cm,TNM stage I〜II and negative ER expression(P<0.05).The/4 values in the positive transfection group at 24,48 and 72 h after culture were(0.487±0.102),(0.702±0.107)and(0.972±0.106),which were significantly higher than those in the control group and the blank transfection group(P<0.05).There was no significant difference in apoptosis rate between the positive transfection group,the blank transfection group and the control group(P>0.05).The number of invasion cells in the positive transfection group was(101.20±11.44),which was significantly higher than that in the blank transfection group and control group(P<0.05).Conclusion CARD9 protein expression in breast cancer tissues is up-regulated,and the up-regulation is related to tumor diameter,TNM stage and ER expression.CARD9 expression exerts certain effect on proliferation and invasion of MDA-MB-231 cells in breast cancer,but has no significant effect on apoptosis.