miR-101对乳腺癌患者的诊断及对乳腺癌细胞自噬和凋亡的影响

The Effect of miR-101 on the Diagnosis of Breast Cancer Patients and Autophagy and Apoptosis of Breast Cancer Cells

目的:探讨血清miR-101在乳腺癌患者中的表达和临床特征,以及对MCF-7细胞自噬和凋亡的影响。方法:选择2021年6月至2022年10月收治的96名乳腺癌患者作为观察组,41例乳腺良性增生患者作为良性组,35名健康体检者作为正常对照组。qRT-PCR检测3组血清miR-101的表达水平;比较乳腺癌患者不同临床病理特征血清miRNA-101表达水平;ROC曲线分析miR-101的诊断价值。应用miR-101 mimic试剂对乳腺癌细胞株(MCF-7)进行转染,qPCR法检测MCF-7和正常乳腺细胞株(HBL-100)miR-101的表达;CCK-8法检测各组细胞的增殖;qPCR法和Western blot法检测细胞自噬和凋亡基因和蛋白的表达;流式细胞术检测各组凋亡率。结果:观察组miRNA-145相对表达量(n=96,0.27)显著低于良性组(n=41,0.63)和对照组(n=35,1.00),差异有统计学意义(P<0.01)。血清miRNA-101的表达在淋巴结转移、分化程度、肿瘤直径和TNM分期差异具有显著性(P<0.01)。ROC曲线分析显示,在3组对比中,曲线下面积分别为0.723(95%CI:0.813-0.924)和0.875(95%CI:0.835~0.981)。CCK-8、qPCR和Western blot结果显示,miR-101过表达可抑制MCF-7细胞增殖,显著增加MCF-7细胞BAX的基因和蛋白表达(P<0.01),降低BCL-2、LC3B和Beclin-1基因和蛋白的表达(P<0.01),增加MCF-7细胞凋亡率。结论:在乳腺癌患者中,血清miR-101的表达水平显著降低,可能成为潜在的乳腺癌的诊断指标。此外,miR-101过表达具有抑制MCF-7乳腺癌细胞增殖、抑制自噬,并促进细胞凋亡的作用。这些结果为进一步研究miR-101在乳腺癌发生和发展中的机制提供了重要线索,为乳腺癌的早期诊断和治疗提供了新的策略。

Objective:To investigate the expression and clinical characteristics of serum miR-101 in patients with breast cancer and its effects on autophagy and apoptosis of MCF-7 cells.Methods:A total of 96 patients with breast cancer treated from June 2021 to October 2022 were selected as the observation group,41 patients with benign breast hyperplasia were selected as the benign group,and 35 healthy subjects were selected as the normal control group.The expression level of miR-101 in serum of the 3 groups was detected by qRT-PCR.The expression levels of serum miRNA-101 in different clinicopathological features of breast cancer patients were compared.The diagnostic value of miR-101 was analyzed by ROC curve.Breast cancer cell line(MCF-7)was transfected with miR-101 mimic reagent.The expression of miR-101 in MCF-7 and normal breast cell line(HBL-100)was detected by qPCR.The cell proliferation was detected by CCK-8 method.The expressions of autophagy and apoptosis genes and proteins were detected by qPCR and Western blot.The apoptosis rate of each group was detected by flow cytometry.Results:The relative expression of miRNA-145 in observation group(n=96,0.27)was significantly lower than that in benign group(n=41,0.63)and control group(n=35,1.00),the difference was statistically significant(P<0.01).The expression of serum miRNA-101 was significantly different in lymph node metastasis,differentiation degree,tumor diameter and TNM stage(P<0.01).ROC curve analysis showed that the area under the curve was 0.723(95%CI:0.813-0.924)and 0.875(95%CI:0.835-0.981)in the three groups of comparison,respectively.The results of CCK-8,qPCR and Western blot showed that over-expression of miR-101 inhibited the proliferation of MCF-7 cells,significantly increased the gene and protein expression of BAX in MCF-7 cells(P<0.01),and decreased the gene and protein expression of BCL-2,LC3B and Beclin-1 in McF-7 cells(P<0.01).The apoptosis rate of MCF-7 cells was increased.Conclusion:In patients with breast cancer,the expression level of serum miR-101 is significantly decreased,which may be a potential diagnostic indicator of breast cancer.In addition,miR-101 overexpression inhibited the proliferation of MCF-7 breast cancer cells,inhibited autophagy,and promoted apoptosis.These results provide important clues for further research on the mechanism of miR-101 in the occurrence and development of breast cancer,and provide new strategies for the early diagnosis and treatment of breast cancer.