广东医学
廣東醫學
엄동의학
GUNAGDONG MEDICAL JOURNAL
2014年
10期
1473-1477
,共5页
张曦%张为民%陈蓓%曾云云
張晞%張為民%陳蓓%曾雲雲
장희%장위민%진배%증운운
非小细胞肺癌%表皮生长因子受体-酪氨酸激酶抑制剂%获得性耐药%上皮-间质转化%胰岛素样生长因子Ⅰ型受体
非小細胞肺癌%錶皮生長因子受體-酪氨痠激酶抑製劑%穫得性耐藥%上皮-間質轉化%胰島素樣生長因子Ⅰ型受體
비소세포폐암%표피생장인자수체-락안산격매억제제%획득성내약%상피-간질전화%이도소양생장인자Ⅰ형수체
non-small lung cancer%epidermal growth factor receptor -tyrosine kinase inhibitors%acquired drug resistance%epithelial-mesenchymal transition%insulin-like growth factor Ⅰreceptor
目的:探讨胰岛素样生长因子Ⅰ型受体( IGF-1R)信号通路及上皮-间质转化( EMT)对EGFR野生型、K-ras突变型非小细胞肺癌A549细胞表皮生长因子酪氨酸激酶抑制剂( EGFR-TKIs)获得性耐药的作用。方法选用EGFR野生型、K-ras突变型的非小细胞肺癌A549细胞,用吉非替尼将其诱导成耐药细胞A549/GR。MTT法检测细胞增殖,划痕实验及Transwell侵袭实验检测细胞侵袭转移能力,qRT-PCR和Western blot 法分别检测EMT标志分子及IGF-1R信号通路相关分子的mRNA和蛋白表达。结果 A549/GR对吉非替尼的敏感性显著下降(P<0.05)。与A549细胞比较,A549/GR细胞形态呈现间质化改变,侵袭和转移能力均显著增加(P<0.05)。 A549/GR细胞中间质标志Vimentin的mRNA和蛋白表达量均显著增多(P<0.05),上皮标志E-Cadherin表达量显著减少(P<0.05)。与A549细胞相比,A549/GR细胞IGF-1R的mRNA和蛋白及其磷酸化表达量均未见明显改变(P>0.05),而AKT磷酸化水平及其mRNA水平明显上调(P<0.05),ERK蛋白及其磷酸化和mRNA表达量均明显增加( P<0.05)。 A549/GR 细胞的 EGFR 及其磷酸化蛋白表达量较 A549细胞明显下降( P<0.05)。结论 EMT及AKT和ERK信号通路的激活均可能在EGFR野生型、K-ras突变型A549细胞的EGFR-TKIs获得性耐药中起重要作用,但获得性耐药与IGF-1R表达无关。
目的:探討胰島素樣生長因子Ⅰ型受體( IGF-1R)信號通路及上皮-間質轉化( EMT)對EGFR野生型、K-ras突變型非小細胞肺癌A549細胞錶皮生長因子酪氨痠激酶抑製劑( EGFR-TKIs)穫得性耐藥的作用。方法選用EGFR野生型、K-ras突變型的非小細胞肺癌A549細胞,用吉非替尼將其誘導成耐藥細胞A549/GR。MTT法檢測細胞增殖,劃痕實驗及Transwell侵襲實驗檢測細胞侵襲轉移能力,qRT-PCR和Western blot 法分彆檢測EMT標誌分子及IGF-1R信號通路相關分子的mRNA和蛋白錶達。結果 A549/GR對吉非替尼的敏感性顯著下降(P<0.05)。與A549細胞比較,A549/GR細胞形態呈現間質化改變,侵襲和轉移能力均顯著增加(P<0.05)。 A549/GR細胞中間質標誌Vimentin的mRNA和蛋白錶達量均顯著增多(P<0.05),上皮標誌E-Cadherin錶達量顯著減少(P<0.05)。與A549細胞相比,A549/GR細胞IGF-1R的mRNA和蛋白及其燐痠化錶達量均未見明顯改變(P>0.05),而AKT燐痠化水平及其mRNA水平明顯上調(P<0.05),ERK蛋白及其燐痠化和mRNA錶達量均明顯增加( P<0.05)。 A549/GR 細胞的 EGFR 及其燐痠化蛋白錶達量較 A549細胞明顯下降( P<0.05)。結論 EMT及AKT和ERK信號通路的激活均可能在EGFR野生型、K-ras突變型A549細胞的EGFR-TKIs穫得性耐藥中起重要作用,但穫得性耐藥與IGF-1R錶達無關。
목적:탐토이도소양생장인자Ⅰ형수체( IGF-1R)신호통로급상피-간질전화( EMT)대EGFR야생형、K-ras돌변형비소세포폐암A549세포표피생장인자락안산격매억제제( EGFR-TKIs)획득성내약적작용。방법선용EGFR야생형、K-ras돌변형적비소세포폐암A549세포,용길비체니장기유도성내약세포A549/GR。MTT법검측세포증식,화흔실험급Transwell침습실험검측세포침습전이능력,qRT-PCR화Western blot 법분별검측EMT표지분자급IGF-1R신호통로상관분자적mRNA화단백표체。결과 A549/GR대길비체니적민감성현저하강(P<0.05)。여A549세포비교,A549/GR세포형태정현간질화개변,침습화전이능력균현저증가(P<0.05)。 A549/GR세포중간질표지Vimentin적mRNA화단백표체량균현저증다(P<0.05),상피표지E-Cadherin표체량현저감소(P<0.05)。여A549세포상비,A549/GR세포IGF-1R적mRNA화단백급기린산화표체량균미견명현개변(P>0.05),이AKT린산화수평급기mRNA수평명현상조(P<0.05),ERK단백급기린산화화mRNA표체량균명현증가( P<0.05)。 A549/GR 세포적 EGFR 급기린산화단백표체량교 A549세포명현하강( P<0.05)。결론 EMT급AKT화ERK신호통로적격활균가능재EGFR야생형、K-ras돌변형A549세포적EGFR-TKIs획득성내약중기중요작용,단획득성내약여IGF-1R표체무관。
Objective To investigate the roles of insulin -like growth factor Ⅰreceptor ( IGF-1R) signal path-way and epithelial-mesenchymal transition ( EMT) in the acquired resistance of epidermal growth factor receptor -tyro-sine kinase inhibitors (EGFR-TKIs) in EGFR wild-type and K-ras mutant non -small cell lung cancer (NSCLC) A549 cells.Methods The EGFR wild-type and K-ras mutant human NSCLC A549 cells were used in this study.The gefitinib-resistant A549 cells (named as A549/GR cells) were created by repeated exposure to gefitinib .MTT assay was used to measure the cell proliferation .Wound-healing array and transwell array were used to determine the invasive and migratory capabilities of cells .The gene and protein expressions of E -cadherin, vimentin, IGF-1R, AKT, ERK and EGFR were determined by qRT -PCR and Western blot, respectively.Results The A549/GR cells acquired resistance to EGFR-TKIs, with significantly reduction in sensitivity to gefitinib (P<0.05).Compared with A549 cells, A549/GR cells presented with mesenchymal phenotype , accompanied by significant enhancement of invasion and migration ( P<0.05).The expression of mesenchymal cell marker vimentin was also significantly increased in A 549/GR cells ( P<0.05), with significant reduction in expression of epithelial cell marker E -Cadherin (P<0.05).No significant differ-ence was found in the expression of IGF -1R mRNA, protein or its phosphorylated form between A 549 and A549/GR cells (P>0.05);while the expression of ERK in A549/GR cells were significantly increased (P<0.05), accompanied with significant up-regulation of mRNA level and phosphorylation level of AKT (P<0.05).The expression of EGFR and its phosphorylated form were significantly reduced in A 549/GR cells as compared with those in A549 cells (P<0.05).Con-clusion EMT and the activation of both AKT and ERK signal pathways may play important roles in acquired EGFR -TKIs-resistance in EGFR wild-type and K-ras mutant NSCLC A549 cells, in which the IGF-1R is not involved .