趋化因子C-C-基元配体5对乳腺癌干细胞增殖和侵袭的影响研究

Effect of C-C motif chemokine ligand 5 on proliferation and invasion of breast cancer stem cells

目的探讨趋化因子C-C-基元配体5(CCL5)对乳腺癌细胞增殖和侵袭能力的影响。方法取在南昌大学第二附属医院行乳腺癌切除术41例患者的乳腺癌组织和癌旁组织作为实验标本,免疫组化法检测将乳腺癌组织和癌旁组织CCL5阳性表达率。将乳腺癌干细胞分为对照组和实验组,对照组不做任何干预,实验组转染抑制CCL5表达质粒(CCL5-siRNA)。用蛋白质印迹法CCL5和Nanog蛋白相对表达量,实时荧光定量逆转录-聚合酶链反应(RT-qPCR)检测转染细胞CCL5 mRNA和Nanog mRNA相对表达量,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法检测转染细胞增殖率,Transwell检测转染细胞侵袭细胞数。结果乳腺患者癌组织和癌旁组织CCL5阳性表达率分别为46.34%和0%,差异有统计学意义(P<0.05)。实验组和对照组CCL5蛋白相对表达量分别为0.57±0.11和1.57±0.26;Nanog蛋白相对表达量分别为1.03±0.12和1.24±0.19;CCL5 mRNA相对表达量分别为0.44±0.08和0.96±0.13;Nanog mRNA相对表达量分别为1.08±0.11和1.18±0.14;细胞增殖率分别为(46.64±7.48)%和(69.32±12.13)%;侵袭细胞数分别为(32.00±5.25)和(48.47±4.22)个,差异均有统计学意义(均P<0.05)。结论乳腺癌组织CCL5表达异常升高,沉默CCL5表达后可抑制乳腺癌干细胞增殖和侵袭能力,其可能作用机制与Nanog基因表达相关。

Objective To investigate the effect of chemokine C-C-ligand 5(CCL5)on the proliferation and invasion of breast cancer cells.Methods Breast cancer tissues and paracancerous tissues of 41 patients who underwent breast cancer resection in the Second Affiliated Hospital of Nanchang University were taken as experimental samples.The positive expression rate of CCL5 in breast cancer tissues and paracancerous tissues was detected by immunohistochemistry.Breast cancer stem cells were divided into control group and test group.The control group was not intervened.The test group was transfected with CCL5 siRNA.The relative expression levels of CCL5 and Nanog proteins were detected by Western blot,the relative expression levels of CCL5 mRNA and Nanog mRNA were detected by quantitative reverse transcription polymerase chain reaction(RT-qPCR),the proliferation rate of transfected cells was detected by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide(MTT),and the number of invasive cells was detected by Transwell.Results The positive expression rates of CCL5 were 46.34%in cancer tissue and 0%in paracancerous tissue,with statistically significant difference(P<0.05).There were statistically significant differences between the test group and the control group in the relative expression of CCL5 protein[0.57±0.11 vs 1.57±0.26],the relative expression of Nanog protein[1.03±0.12 vs 1.24±0.19],the relative expression of CCL5 mR NA[0.44±0.08 vs 0.96±0.13],the relative expression of Nanog mR NA[1.08±0.11 vs 1.18±0.14],the cell proliferation rate[(46.64±7.48)%vs(69.32±12.13)%],and the number of invasive cells[32.00±5.25 vs 48.47±4.22](all P<0.05).Conclusion The expression of CCL5 in breast cancer is abnormally high.Silencing CCL5 expression can inhibit the proliferation and invasion of breast cancer stem cells,and the possible mechanism is related to the expression of Nanog gene.