FRY蛋白在乳腺癌中的表达和临床意义及对癌细胞增殖、迁移和凋亡能力的影响

The research of the expression and clinical significance of FRY protein in breast cancer and its effect on proliferation,migration and apoptosis of cancer cells

目的:探讨FRY蛋白在乳腺癌中的表达和临床意义及对癌细胞增殖、迁移和凋亡能力的影响。方法:收集2017年6月-2018年6月于我院接受手术治疗的87例乳腺癌患者癌组织及其距离癌组织超过2.5 cm处的癌旁组织,另收集同期因良性乳腺肿瘤接受手术的34例患者的病变组织,免疫组化检测FRY蛋白的表达情况。将FRY的表达与乳腺癌的相关临床病理指标进行相关性分析。构建rAAV9-FRY过表达载体和对照rAAV9-FRY-NC,转染人乳腺癌细胞株MCF7,实验分为正常组(细胞常规培养,不进行干预)、对照组(细胞转染rAAV9-FRY-NC后,常规培养)、实验组(细胞转染rAAV9-FRY后,常规培养);EdU标记实验、吖啶橙(acridine orange,AO)/溴化乙锭(ethidium bromide,EB)双染法、Transwell小室、裸鼠肿瘤异种移植模型、Western blot检测各组MCF细胞的增殖、凋亡、迁移、在裸鼠体内增殖与肺转移的情况、FRY和神经纤毛蛋白-1(neuropilin-1,NRP1)表达的水平。结果:免疫组化结果显示,FRY在癌旁组织中阳性率最高,FRY的阳性表达在良性肿瘤中明显高于恶性乳腺癌组织(P <0.05)。FRY的阳性表达在乳腺癌的不同分级具有统计学差异(P <0.05),不同年龄以及TNM分期的FRY表达比较差异无统计学意义(P> 0.05);而肿瘤直径和浸润深度与FRY的阳性表达相关(P <0.05)。与正常组和对照组相比,实验组MCF7细胞的EdU阳性细胞比例、迁移能力、移植瘤的瘤体重量、瘤体肺脏转移的病灶比例及NRP1的表达明显下降;细胞凋亡率、FRY的表达明显上升;差异均具有统计学意义(P <0.05)。结论:FRY在乳腺癌组织中表达下调,在乳腺癌细胞中FRY特异性结合NRP1来抑制细胞的增殖、迁移,并促进其凋亡。

Objective:To investigate the expression and clinical significance of FRY protein in breast cancer and its effect on the proliferation,migration and apoptosis of cancer cells.Methods:From June 2017 to June 2018,the cancer tissues of 87 cases of breast cancer patients who received surgical treatment in our hospital and their adjacent tissues more than 2.5 cm away from the cancer tissues were collected.In addition,the pathological tissues of 34 cases of patients who received surgery for benign breast tumors during the same period were collected,and the expression of FRY protein was detected by immunohistochemistry.Correlation analysis was performed between the expression of FRY and clinical pathological parameters of breast cancer.The rAAV9-FRY overexpression vector and the control rAAV9-FRY-NC were constructed and transfected into human breast cancer cell line MCF7.The experiment was divided into normal group(normal cell culture,without any intervention),control group(cell transfection rAAV9-FRY-NC,routine culture),experimental group(cell transfection of rAAV9-FRY,routine culture).EdU labeling,acridine orange(AO)/ethidium bromide(EB) double staining,Transwell cab,nude mouse tumor xenograft model,Western blot analysis were used to detect the proliferation,apoptosis,migration,proliferation and lung metastasis in nude mice,the level of expression FRY and neuropilin-1(NRP1) in each group.Results:Immunohistochemical results showed that the positive rate was the highest in the adjacent tissues,and the positive expression of FRY was significantly higher in benign tumors than in malignant breast cancer tissues(P<0.05).The positive expression of FRY was statistically significant in different grades of breast cancer(P<0.05).There was no significant difference in FRY expression in different ages and TNM stages(P>0.05).Tumor size and the depth of invasion were correlated with positive expression of FRY(P<0.05).Compared with the normal group and the control group,the proportion of EdU-positive cells,the migration ability,the tumor weight of the transplanted tumor,the proportion of the tumor metastasis of the tumor and the expression of NRP1 were significantly decreased in the experimental group.The cell apoptosis rate and the expression of FRY increased significantly.The differences were statistically significant(P<0.05).Conclusion:FRY was down-regulated in breast cancer tissues,and FRY specifically combined with NRP1 in breast cancer cells to inhibit cell proliferation,migration and promote apoptosis.