摘要
目的研究氨基酸转运蛋白SLC6A14在大肠癌中的表达特征及其促进大肠癌细胞生长的作用。方法采用实时定量荧光PCR(real-time PCR)检测35对大肠癌及癌旁组织中SLC6A14 mRNA的表达水平;Western印迹法检测SLC6A14在大肠癌细胞株HT-29和SW620中的蛋白表达;免疫组化法检测108例大肠癌组织中SLC6A14的蛋白表达并分析其表达强度与患者临床特征的相关性;MTT法和流式细胞术分别检测下调或抑制SLC6A14对大肠癌细胞生长及凋亡的影响。结果SLC6A14 mRNA在大肠癌组织中的表达水平是癌旁组织的(3.98±1.1)倍(P<0.05);SLC6A14在大肠癌细胞株HT-29和SW620中的蛋白表达明显高于肝癌细胞株Huh7,分别是其(4.31±1.46)倍和(3.14±0.93)倍,均P<0.05。免疫组化结果显示SLC6A14在Ⅳ期癌组织中的蛋白表达强度(135.36±24.28)明显高于Ⅰ、Ⅱ、Ⅲ期癌组织(分别为42.87±26.50、75.67±30.25、101.32±32.47,均P<0.001),但其表达水平在不同年龄、性别和肿瘤大小中的表达差异未见统计学意义(P>0.05)。通过siRNA下调SLC6A14在HT-29和SW620中的表达或抑制剂α-MT后,MTT结果显示大肠癌细胞增殖明显减慢,流式细胞术检测Annexin-Ⅴ+/PI-细胞凋亡明显增加,P<0.05。结论SLC6A14在大肠癌组织和细胞株中表达增加并促进癌细胞生长,是大肠癌治疗的潜在靶点。
Objective To investigate the expression of amino acid transporter SLC6A14 and its effect on cell proliferation in colorectal cancer(CRC).Methods The expression of SLC6A14 mRNA in 35 paired colorectal cancer and pericancerous tissue samples was detected by real-time RT-PCR(qRT-PCR).The expression of SLC6A14 protein in tissue samples of 108 CRC patients was determined by immunohistochemistry,and the correlation between the intensity of SLC6A14 expression and the clinical characteristics of patients was analyzed.The expression of SLC6A14 protein in human colorectal cancer HT-29,SW620 cells and human liver cancer Huh7 cells was detected by Western blotting.HT-29 and SW620 cells were treated with siRNA orα-MT to down-regulating SLC6A14 expression;and the cell proliferation and apoptosis were detected by MTT method and flow cytometry.Results The expression of SLC6A14 mRNA in colorectal cancer tissues was 3.98±1.1 times higher than that in pericancerous tissues(P<0.05).The expression of SLC6A14 protein in HT-29 and SW620 cells was 4.31±1.46 and 3.14±0.93 times higher than that in Huh7 cells(P<0.05).Immunohistochemical results showed that the expression intensity of SLC6A14 in stageⅣof CRC(135.36±24.28)was significantly higher than that in stageⅠ,ⅡandⅢ(42.87±26.50,75.67±30.25,101.32±32.47,respectively)(P<0.001),but there was no significant difference in the expression level of SLC6A14 among patients with different age,gender and tumor size(P>0.05).After HT-29 and SW620 cells were treated with SLC6A14-siRNA or inhibitorα-MT at three different concentrations,the cell proliferation was significantly reduced,and the apoptosis was significantly increased(P<0.05).Conclusion SLC6A14 is upregulated in colorectal cancer and is involved in proliferation of CRC cells,indicating that it may be a potential therapeutic target for CRC.
作者
罗轶玮
符雪莲
马钻
王恺京
李芳漪
陆英
LUO Yi-wei;FU Xue-lian;MA Zuan;WANG Kai-jing;LI Fang-yi;LU Ying(Dept.of General Practice,East Hospital,Tongji University School of Medicine,Shanghai 200120,China;Research Center for Translational Medicine,East Hospital,Tongji University School of Medicine,Shanghai 200120,China)
出处
《同济大学学报（医学版）》
CAS
2021年第1期28-34,共7页
Journal of Tongji University(Medical Science)
基金
上海市浦东新区科委创新基金(PKJ2016-Y63)
国家自然科学基金青年项目(82001725)
江西省自然科学基金面上项目(20181BAB205033)。
