中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2013年
10期
690-695
,共6页
徐莹%王舒蓓%王海郦%孙蕴伟
徐瑩%王舒蓓%王海酈%孫蘊偉
서형%왕서배%왕해역%손온위
上皮细胞%结肠肿瘤%微RNAs%喹唑啉类%钙黏着糖蛋白类%细胞系,肿瘤
上皮細胞%結腸腫瘤%微RNAs%喹唑啉類%鈣黏著糖蛋白類%細胞繫,腫瘤
상피세포%결장종류%미RNAs%규서람류%개점착당단백류%세포계,종류
Epithelial cells%Colonic neoplasms%MicroRNAs%Quinazolines%Cadherins%Cell Line,tumor
目的 探讨微小核糖核酸(miRNA)-200c及上皮间质转化(EMT)与结肠癌细胞对吉非替尼治疗敏感性的关系.方法 细胞计数试剂盒8(CCK-8)法检测吉非替尼对4种人结肠癌细胞系HT29、SW620、HCT116、SW480的生长抑制作用；以实时定量PCR法检测4种结肠癌细胞中miRNA-200c,上皮标志物(E-钙黏蛋白),间质标志物[波形蛋白、具锌指E盒结构的同源异性盒1(ZEB 1)]mRNA水平表达,Western印迹检测E-钙黏蛋白、波形蛋白、ZEB 1蛋白水平表达.外源性上调或下调miRNA-200c表达,观察EMT相关基因表达变化及细胞对吉非替尼敏感性的变化.结果 HT29细胞对吉非替尼最为敏感[半数抑制浓度(IC50)=(7.70±0.31) μmol/I],其miRNA-200c与E-钙黏蛋白表达量均为最高,波形蛋白及ZEB 1表达水平极低；HCT116及SW480细胞系对吉非替尼为中度敏感[IC50=(11.88±0.97),(16.63士0.45)μmol/L],其miRNA-200c、E-钙黏蛋白呈中等程度表达；SW620细胞系对吉非替尼最不敏感[IC50=(26.43±3.68) μmol/L],miRNA-200c与E-钙黏蛋白表达量最低,波形蛋白及ZEB 1表达量均显著高于其他3种细胞系.外源性上调miRNA-200c表达后,SW620细胞中E-钙黏蛋白表达上调,ZEB 1及波形蛋白表达下调,同时细胞对吉非替尼的敏感性亦显著提高；相反,外源性下调miRNA-200c表达后,HT29细胞中E-钙黏蛋白表达下调,ZEB 1及波形蛋白表达上调,同时细胞对吉非替尼的敏感性显著降低.结论 miRNA-200c可能通过调控EMT,上调E-钙黏蛋白表达,进而影响结肠癌细胞对吉非替尼的敏感性.
目的 探討微小覈糖覈痠(miRNA)-200c及上皮間質轉化(EMT)與結腸癌細胞對吉非替尼治療敏感性的關繫.方法 細胞計數試劑盒8(CCK-8)法檢測吉非替尼對4種人結腸癌細胞繫HT29、SW620、HCT116、SW480的生長抑製作用；以實時定量PCR法檢測4種結腸癌細胞中miRNA-200c,上皮標誌物(E-鈣黏蛋白),間質標誌物[波形蛋白、具鋅指E盒結構的同源異性盒1(ZEB 1)]mRNA水平錶達,Western印跡檢測E-鈣黏蛋白、波形蛋白、ZEB 1蛋白水平錶達.外源性上調或下調miRNA-200c錶達,觀察EMT相關基因錶達變化及細胞對吉非替尼敏感性的變化.結果 HT29細胞對吉非替尼最為敏感[半數抑製濃度(IC50)=(7.70±0.31) μmol/I],其miRNA-200c與E-鈣黏蛋白錶達量均為最高,波形蛋白及ZEB 1錶達水平極低；HCT116及SW480細胞繫對吉非替尼為中度敏感[IC50=(11.88±0.97),(16.63士0.45)μmol/L],其miRNA-200c、E-鈣黏蛋白呈中等程度錶達；SW620細胞繫對吉非替尼最不敏感[IC50=(26.43±3.68) μmol/L],miRNA-200c與E-鈣黏蛋白錶達量最低,波形蛋白及ZEB 1錶達量均顯著高于其他3種細胞繫.外源性上調miRNA-200c錶達後,SW620細胞中E-鈣黏蛋白錶達上調,ZEB 1及波形蛋白錶達下調,同時細胞對吉非替尼的敏感性亦顯著提高；相反,外源性下調miRNA-200c錶達後,HT29細胞中E-鈣黏蛋白錶達下調,ZEB 1及波形蛋白錶達上調,同時細胞對吉非替尼的敏感性顯著降低.結論 miRNA-200c可能通過調控EMT,上調E-鈣黏蛋白錶達,進而影響結腸癌細胞對吉非替尼的敏感性.
목적 탐토미소핵당핵산(miRNA)-200c급상피간질전화(EMT)여결장암세포대길비체니치료민감성적관계.방법 세포계수시제합8(CCK-8)법검측길비체니대4충인결장암세포계HT29、SW620、HCT116、SW480적생장억제작용；이실시정량PCR법검측4충결장암세포중miRNA-200c,상피표지물(E-개점단백),간질표지물[파형단백、구자지E합결구적동원이성합1(ZEB 1)]mRNA수평표체,Western인적검측E-개점단백、파형단백、ZEB 1단백수평표체.외원성상조혹하조miRNA-200c표체,관찰EMT상관기인표체변화급세포대길비체니민감성적변화.결과 HT29세포대길비체니최위민감[반수억제농도(IC50)=(7.70±0.31) μmol/I],기miRNA-200c여E-개점단백표체량균위최고,파형단백급ZEB 1표체수평겁저；HCT116급SW480세포계대길비체니위중도민감[IC50=(11.88±0.97),(16.63사0.45)μmol/L],기miRNA-200c、E-개점단백정중등정도표체；SW620세포계대길비체니최불민감[IC50=(26.43±3.68) μmol/L],miRNA-200c여E-개점단백표체량최저,파형단백급ZEB 1표체량균현저고우기타3충세포계.외원성상조miRNA-200c표체후,SW620세포중E-개점단백표체상조,ZEB 1급파형단백표체하조,동시세포대길비체니적민감성역현저제고；상반,외원성하조miRNA-200c표체후,HT29세포중E-개점단백표체하조,ZEB 1급파형단백표체상조,동시세포대길비체니적민감성현저강저.결론 miRNA-200c가능통과조공EMT,상조E-개점단백표체,진이영향결장암세포대길비체니적민감성.
Objective To explore the relationship between microRNA (miRNA)-200c expression,epithelial-mesenchymal transition (EMT) and the sensitivity to gefitinib in colon cancer cells.Methods The inhibitory effects of gefitinib on four types of colon cancer cell lines (HT29,SW620,HCT1116,SW480) were examined by cell counting kit-8 (CCK-8) assay.The expressions of miRNA-200c,epithelial marker (E-cadherin) and mesenchymal markers (vimentin and zinc finger Ebox binding homeobox 1 ZEB 1) at mRNA level in four types of colon cancer cell lines were detected by fluorescence quantitative polymerase chain reaction.The expressions of E-cadherin,vimentin and ZEB 1 at protein level were determined by Western blot.After up-or down-regulated the expression of miRNA-200c,the changes of the expression of EMT related genes and the sensitivity to gefitinib were observed.Results HT29 cells were most sensitive to gefitinib (IC50 =(7.70 ± 0.31) μmol/L),in which the expressions of both miRNA-200c and E-cadherin were the highest,and the expressions of vimentin and ZEB1 were extremely low.HCT116 and SW480 were moderately sensitive to gefitinib (IC50=(11.88±0.97) and (16.63±0.45) μmol/L),and the expressions of miRNA-200c and Ecadherin were moderate.SW620 cell line was most insensitive to gefitinib (IC50 =(26.43 ± 3.68)μmol/L),the expressions of miRNA-200c and E-cadherin were the lowest.The expressions of vimentin and ZEB 1 in SW620 were higher than that of the other three types of cell lines.After upregulated the expression of miRNA 200c,the expression of E-cadherin in SW620 cells increased,the expression of ZEB 1 and vimentin decreased,and the sensitivity to gefitinib increased.After downregulated the expression of miRNA-200c,the expression of E-cadherin in HT29 cells decreased,the expression of ZEB 1 and vimentin increased,and the sensitivity to gefitinib decreased.Conclusion miRNA-200c may up-regulate the expression of E-cadherin through EMT regulation,and then influence the sensitivity to gefitinib in colon cancer cells.
