论文部分内容阅读
目的:探讨PH结构域且富含亮氨酸重复基序的丝/苏氨酸蛋白磷酸酶1(PHLPP1)对乳腺癌恶性细胞行为的影响及其机制。方法:采用蛋白质免疫印迹法(Western blot)检测河南省人民医院2017年6月到2019年6月手术切除的59对乳腺癌组织和配对癌旁组织的PHLPP1蛋白表达水平。采用慢病毒在MDA-MB-231乳腺癌细胞系构建对照和PHLPP1过表达稳定细胞系，分别命名为对照组和实验组。采用细胞计数试剂盒(CCK-8)测定两组细胞增殖能力；采用凋亡试剂盒检测两组细胞凋亡水平；采用Transwell分析两组细胞侵袭能力；采用异体移植瘤实验分析细胞在体内生长和转移能力；采用Western blot分析两组基质金属蛋白酶3(MMP-3)和半胱氨酸天冬氨酸蛋白酶(Caspase)-3蛋白表达水平。两组间比较行独立样本的 n t检验。n 结果:癌旁组织中PHLPP1蛋白相对表达水平(1.54±0.23)明显高于乳腺癌组织 (0.74±0.15)，差异有统计学意义(n t＝3.749，n P<0.05)。对照组细胞吸光度(n A)值(1.98±0.22)明显高于观察组细胞n A值(1.06±0.16)，差异有统计学意义(n t＝4.082，n P<0.05)。对照组细胞体内生长30 d后肿瘤体积[(1 209.43±289.69) mmn 3]明显高于观察组细胞[(920.43±100.54) mmn 3]，差异有统计学意义(n t＝6.943，n P<0.05)。对照组细胞凋亡比例[(4.19±4.90)%]明显低于观察组细胞凋亡比例[(27.33±4.98)%]，差异有统计学意义(n t＝3.431，n P<0.05)。对照组细胞迁移数量[(91.49±7.23)个]明显高于观察组细胞迁移数量[(57.18±6.20)个]，差异有统计学意义(n t＝5.298，n P<0.05)。对照组细胞体内生长30 d后淋巴结转移数量[(19.48±3.99)个]明显高于观察组细胞[(7.19±1.54)个]，差异有统计学意义(n t＝6.943，n P<0.05)。对照组细胞Caspase-3蛋白相对表达水平(0.86±0.18)明显低于观察组细胞(1.59±0.25)，差异有统计学意义(n t＝3.002，n P<0.05)。对照组细胞MMP-3蛋白相对表达水平(1.32±0.20)明显高于观察组细胞(0.73±0.17)，差异有统计学意义(n t＝3.691，n P<0.05)。n 结论:PHLPP1在乳腺癌组织中呈低表达，上调PHLPP1表达水平促进细胞凋亡，抑制细胞生长、侵袭和转移。“,”Objective:To investigate the effect of PH domain and leucine rich serine/threonine protein phosphatase 1 (PHLPP1) on the behaviors of breast cancer cells and the underlying molecular mechanism.Methods:The expression of PHLPP1 protein in 59 pairs of breast cancer tissues and adjacent tissues from June 2017 to June 2019 was detected by Western blotting. Lentivirus was used to construct control and PHLPP1 overexpression stable cell lines in MDA-MB-231 breast cancer cell line, named control group and experimental group respectively. The cell proliferation of the two groups was analyzed by cell counting kit-8 (CCK-8). The cell apoptosis level of the two groups was analyzed by apoptosis kit. The invasion ability of the two groups was analyzed by Transwell. The growth and metastasis ability of the cells n in vivo was analyzed by xenograft tumor experiment, and the protein expression levels of matrix metalloproteinase-3 (MMP-3) and cysteinyl aspartate-specific protease (Caspase)-3 were analyzed by Western blotting. SPSS 17.0 statistical software was used to analyze the data and the data were expressed as mean±standard deviation. The comparison between two groups was performed by n t test of independent samples.n Results:The relative expression level of PHLPP1 protein in paracancerous tissues (1.54±0.23) was significantly higher than that in breast cancer tissues (0.74±0.15, n t=3.749, n P<0.05). The absorbance value in control group (1.98±0.22) was significantly higher than that in observation group (1.06±0.16) (n t=4.082, n P<0.05). The tumor volume in the control group [(1 209.43±289.69) mmn 3] was significantly greater than that in the observation group [(920.43±100.54) mmn 3 ] (n t=6.943, n P<0.05). The percentage of apoptosis in the control group [(4.19±4.90)%] was significantly higher than that in the observation group [(27.33±4.98)%] (n t=3.431, n P<0.05). The number of migrating cells in the control group (91.49±7.23) was significantly greater than that in the observation group (57.18±6.20,n t=5.298, n P<0.05). The number of lymph node metastasis in the control group was significantly greater than that in the observation group (19.48±3.99 vs. 7.19±1.54,n t=6.943, n P<0.05). The relative expression level of Caspase-3 protein in the control group (0.86±0.18) was significantly lower than that in the observation group (1.59±0.25,n t=3.002, n P<0.05). The relative expression level of MMP-3 protein in the control group (1.32±0.20) was significantly higher than that in the observation group (0.73±0.17,n t=3.691, n P<0.05).n Conclusion:The expression of PHLPP1 is low in breast cancer tissues. Up-regulating the expression of PHLPP1 can promote apoptosis and inhibit cell growth, invasion and metastasis.