性别决定相关基因簇9对胰腺癌干细胞增殖、侵袭的影响及其与吉西他滨耐药性的关系  

Effects of sex determining region Y box 9 on proliferation and invasion of pancreatic cancer stem cells and its relationship with gemcitabine resistance

作　　者：沈俊 周锐 王建祥 Shen Jun;Zhou Rui;Wang Jianxiang(Department of General Surgery,Puai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430033,China)

机构地区：[1]华中科技大学同济医学院附属普爱医院普外科,武汉430033

出　　处：《中华实验外科杂志》2020年第7期1255-1258,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨性别决定相关基因簇9(SOX9)对胰腺癌干细胞增殖、侵袭的影响及其与吉西他滨耐药性的关系。方法流式细胞术分选PANC-1细胞(购自中国科学院上海细胞生物学研究所)中以CD24+CD44+ESA+为标志物的细胞亚群,并通过小鼠(购自北京维通利华实验动物技术有限公司)移植成瘤实验验证胰腺癌干细胞的特性。分选的胰腺癌干细胞分为对照组、shNC组和shSOX9组。实时定量反转录聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)检测各组细胞中SOX9的表达。细胞计数试剂盒(CCK-8)检测各组细胞的增殖能力。Transwell细胞侵袭实验检测各组细胞的侵袭能力。CCK-8检测各组细胞对吉西他滨的敏感性。细胞流式术检测各组细胞经吉西他滨处理后的凋亡。多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验,率的比较采用χ2检验。结果 PANC-1细胞中分选出CD24+CD44+ESA+细胞(占0.8%),其在小鼠皮下移植后的移植瘤体积[(567.35±23.21) mm3]大于接种PANC-1细胞小鼠的皮下移植瘤体积[(106.83±12.43) mm3,t=18.040,P<0.01]。SOX9 mRNA在shSOX9组中的相对表达量(0.42±0.05)低于对照组(1.00±0.03)和shNC组(1.01±0.03,F=77.470,P<0.01);SOX9蛋白表达量在shSOX9组(0.12±0.04)中低于对照组(0.64±0.12)和shNC组(0.62±0.07,F=85.230,P<0.01)。shSOX9组细胞的吸光度值(0.74±0.13)低于对照组(1.38±0.21)和shNC组(1.37±0.23,F=78.320,P<0.01)。shSOX9组侵袭细胞数目[(162.20±16.34)个]低于对照组[(250.50±24.21)个]和shNC组[(253.13±21.78)个,F=21.040,P<0.01]。shSOX9组吉西他滨的半数抑制浓度(IC50)值为(2.78±0.08) mg/L,低于对照组[(5.12±0.13) mg/L]和shNC组[(5.08±0.11) mg/L,F=23.450,P<0.01]。shSOX9组细胞的凋亡率为[(35.93±3.25)%],高于对照组[(20.08±3.14)%]和shNC组[(19.76±2.67)%,χ2=50.130,P<0.01]。结论沉默SOX9可抑制胰腺癌干细胞增殖与侵袭,提高胰腺癌干细胞对吉西他滨的敏感性。Objective:To explore the effect of sex determining region Y box 9(SOX9)on the proliferation and invasion of pancreatic cancer stem cells and its relationship with gemcitabine resistance.Methods:Flow cytometry was used to sort PANC-1 cells with CD24+CD44+ESA+as a marker,and the characteristics of pancreatic cancer stem cells were verified by tumor transplantation experiments in mice.The sorted pancreatic cancer stem cells were divided into control group,shNC group and shSOX9 group.Real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the expression of SOX9 in each group.Cell counting kit-8(CCK-8)was used to detect the proliferation ability of cells in each group.Transwell cell invasion experiment was used to detect the invasion ability of cells in each group.CCK-8 was used to detect the sensitivity of cells to gemcitabine in each group.Flow cytometry was used to detect the apoptosis of cells in each group after treatment with gemcitabine.Results:CD24+CD44+ESA+cells(0.8%)were selected from PANC-1 cells,and the volume of transplanted tumors after subcutaneous transplantation in mice[(567.35±23.21)mm 3]was larger than that of mice inoculated with PANC-1 cells[(106.83±12.43)mm 3,t=18.040,P<0.01].The relative expression of SOX9 mRNA in shSOX9 group(0.42±0.05)was significantly lower than that in control group(1.00±0.03)and shNC group(1.01±0.03)(F=77.470,P<0.01).The expression level of SOX9 protein in shSOX9 group(0.12±0.04)was significantly lower than that in control group(0.64±0.12)and shNC group 0.62±0.07)(F=85.230,P<0.01).The absorbance value of cells in shSOX9 group(0.74±0.13)was significantly lower than that in control group(1.38±0.21)and shNC group(1.37±0.23)(F=78.320,P<0.01).The number of invasive cells in shSOX9 group[(162.20±16.34)cells]was significantly lower than that in control group[(250.50±24.21)cells]and shNC group[(253.13±21.78)cells,F=21.040,P<0.01].The IC 50 value of gemcitabine in the shSOX9 group was[(2.78±0.08)mg/L],whi

关 键 词：胰腺癌 性别决定相关基因簇9 胰腺癌干细胞 吉西他滨 

分 类 号：R735.9[医药卫生—肿瘤]