中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
7期
1324-1327
,共4页
庞明辉%王康%伍刚%梁耀泽%胡阳%何飞%侯能易%李平%李国新
龐明輝%王康%伍剛%樑耀澤%鬍暘%何飛%侯能易%李平%李國新
방명휘%왕강%오강%량요택%호양%하비%후능역%리평%리국신
Lin28b%结肠癌%奥沙利铂%化疗
Lin28b%結腸癌%奧沙利鉑%化療
Lin28b%결장암%오사리박%화료
Lin28b%Colon carcinoma%Oxaliplatin%Chemosensitivity
目的 探讨人结肠癌细胞中Lin28b基因表达及其与奥沙利铂化疗敏感性之间的关系.方法 构建干扰Lin28b的shRNA质粒(sh-Lin28b),并转染至结肠癌细胞(Cac02、SW480和HCT116细胞)；逆转录-聚合酶链反应(RT-PCR)和Western blot法分别检测结肠癌细胞中Lin28b在mRNA和蛋白水平的表达；用细胞计数试剂盒(CCK-8)检测sh-Lin28b和奥沙利铂化疗协同作用,Transwell实验检测处理后的结肠癌细胞迁移能力.结果 RT-PCR和Western blot结果证实Cac02、HCT116及SW480结肠癌细胞中均有Lin28b基因表达,其中在SW480细胞中的蛋白表达量比在HCT116中高1.83倍.SW480和HCT116细胞中Lin28b的mRNA表达在50 nmol/L的sh-Lin28b作用下分别被下调36.4％和90.2％,在100 nmol/L的sh-Lin28b的条件下分别被下调61.8％和89.5％.CCK-8结果显示奥沙利铂处理48 h后,HCT116细胞的IC50值为(6.09±1.42) mg/L,SW480细胞升高34.3％,至(8.18 ±3.64) mg/L.sh-Lin28b联合奥沙利铂处理组比较于对照转染组(NC)抑制SW480和HCTll6细胞生存率分别为(55.10±0.78)％和(50.30±0.69)％.Transwell 实验结果显示SW480细胞的迁移能力在sh-Lin28b的作用下降低至(48.60±0.92)％.结论 沉默表达于结肠癌细胞的Lin28b,可显著抑制结肠癌细胞的迁移能力,并促进奥沙利铂的化疗敏感性.
目的 探討人結腸癌細胞中Lin28b基因錶達及其與奧沙利鉑化療敏感性之間的關繫.方法 構建榦擾Lin28b的shRNA質粒(sh-Lin28b),併轉染至結腸癌細胞(Cac02、SW480和HCT116細胞)；逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot法分彆檢測結腸癌細胞中Lin28b在mRNA和蛋白水平的錶達；用細胞計數試劑盒(CCK-8)檢測sh-Lin28b和奧沙利鉑化療協同作用,Transwell實驗檢測處理後的結腸癌細胞遷移能力.結果 RT-PCR和Western blot結果證實Cac02、HCT116及SW480結腸癌細胞中均有Lin28b基因錶達,其中在SW480細胞中的蛋白錶達量比在HCT116中高1.83倍.SW480和HCT116細胞中Lin28b的mRNA錶達在50 nmol/L的sh-Lin28b作用下分彆被下調36.4％和90.2％,在100 nmol/L的sh-Lin28b的條件下分彆被下調61.8％和89.5％.CCK-8結果顯示奧沙利鉑處理48 h後,HCT116細胞的IC50值為(6.09±1.42) mg/L,SW480細胞升高34.3％,至(8.18 ±3.64) mg/L.sh-Lin28b聯閤奧沙利鉑處理組比較于對照轉染組(NC)抑製SW480和HCTll6細胞生存率分彆為(55.10±0.78)％和(50.30±0.69)％.Transwell 實驗結果顯示SW480細胞的遷移能力在sh-Lin28b的作用下降低至(48.60±0.92)％.結論 沉默錶達于結腸癌細胞的Lin28b,可顯著抑製結腸癌細胞的遷移能力,併促進奧沙利鉑的化療敏感性.
목적 탐토인결장암세포중Lin28b기인표체급기여오사리박화료민감성지간적관계.방법 구건간우Lin28b적shRNA질립(sh-Lin28b),병전염지결장암세포(Cac02、SW480화HCT116세포)；역전록-취합매련반응(RT-PCR)화Western blot법분별검측결장암세포중Lin28b재mRNA화단백수평적표체；용세포계수시제합(CCK-8)검측sh-Lin28b화오사리박화료협동작용,Transwell실험검측처리후적결장암세포천이능력.결과 RT-PCR화Western blot결과증실Cac02、HCT116급SW480결장암세포중균유Lin28b기인표체,기중재SW480세포중적단백표체량비재HCT116중고1.83배.SW480화HCT116세포중Lin28b적mRNA표체재50 nmol/L적sh-Lin28b작용하분별피하조36.4％화90.2％,재100 nmol/L적sh-Lin28b적조건하분별피하조61.8％화89.5％.CCK-8결과현시오사리박처리48 h후,HCT116세포적IC50치위(6.09±1.42) mg/L,SW480세포승고34.3％,지(8.18 ±3.64) mg/L.sh-Lin28b연합오사리박처리조비교우대조전염조(NC)억제SW480화HCTll6세포생존솔분별위(55.10±0.78)％화(50.30±0.69)％.Transwell 실험결과현시SW480세포적천이능력재sh-Lin28b적작용하강저지(48.60±0.92)％.결론 침묵표체우결장암세포적Lin28b,가현저억제결장암세포적천이능력,병촉진오사리박적화료민감성.
Objective To investigate the effects of Lin28b expression on the chemosensitivity of colon cancer cells to oxaliplatin.Methods Basic Lin28b mRNA and protein expression level was examined in colon cancer cells by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting,respectively.The half maximal inhibitory concentration of colon cancer cells ( IC50 ) tranfected with shLin28b and treated by oxaliplatin was calculated using cell counting Kit-8 ( CCK-8 ) technique.The migration of theses cells was determined with Transwell method to explore the synergistic effect of sh-Lin28b expression and oxaliplatin.Results There was Lin28b expression in Caco2,HCT116 and SW480 colon cancer cell lines detected by RT-PCR and Western blotting.The protein expression level of Lin28b in SW480 cells was 1.83 times higher than that in HCT116 cells.The treatment of 50 nmol/L sh-Lin28b resulted in 36.4％ and 90.2％ reduction of mRNA expression level of Lin28b in SW480 cells and HCT116 cells,respectively,and that of 100 nmol/L sh-Lin28b led to 61.8％ and 89.5％ reduction,respectively.CCK-8 results showed that the IC50 (6.09 ± 1.42 ) mg/L of HCT116 cells treated with oxaliplatin by 48 h was increased to 34.3％ (8.18 ±3.64) mg/L in SW480 cells.The half maximal inhibitory concentration of transfected SW480 and HCT116 cells was decreased to (55.10 ± 0.78 ) ％ and ( 50.30 -± 0.69 ) ％ by treatment with oxaliplatin as compared with that in normal control group.The Transwell results showed that the migration of transfected SW480 cells was reduced to ( 48.60 ± 0.92 ) ％.Conclusion Silencing the expression of Lin28b in colon cancer cells could greatly inhibit their migration and promote their chemosensitivity to oxaliplatin.
