中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2013年
7期
455-459
,共5页
方霞%顾盼%周彩存%任胜祥%罗本芳%曾郁%吴运瑾%赵印敏%朱旭友
方霞%顧盼%週綵存%任勝祥%囉本芳%曾鬱%吳運瑾%趙印敏%硃旭友
방하%고반%주채존%임성상%라본방%증욱%오운근%조인민%주욱우
癌,非小细胞肺%受体,表皮生长因子%蛋白酪氨酸激酶类%β连环素
癌,非小細胞肺%受體,錶皮生長因子%蛋白酪氨痠激酶類%β連環素
암,비소세포폐%수체,표피생장인자%단백락안산격매류%β련배소
Carcinoma,non-small-cell lung%Receptor,epidermal growth factor%Protein-tyrosine kinases%Beta catenin
目的 探讨下调Wnt通路活性对吉非替尼抑制非小细胞肺癌细胞株增殖的影响及其潜在的作用机制.方法 不同浓度吉非替尼处理PC9细胞株(吉非替尼敏感株)与PC9/AB2细胞株(吉非替尼耐药株),细胞计数检测试剂盒(CCK8)检测两种细胞株的增殖情况.Western blot分别检测PC9与PC9/AB2细胞中Wnt信号通路相关蛋白的表达情况.双荧光素酶报告基因系统(TOP Flash)分别检测PC9和PC9/AB2细胞株Wnt通路的转录活性.采用β-catenin序列特异的siRNA抑制PC9/AB2中Wnt通路的转录活性后,用不同浓度吉非替尼处理,CCK8检测干预后细胞增殖情况.干扰后表皮生长因子受体及其下游通路的表达情况.结果 不同浓度吉非替尼作用下,PC9/AB2细胞对吉非替尼的耐药性明显增高(P＜0.05).PC9/AB2细胞Wnt通路相关蛋白的表达明显高于PC9,差异有统计学意义(t=24.590,P=0.000).TOP Flash结果显示,PC9/AB2细胞株中Wnt通路的转录活性高于PC9细胞株,差异有统计学意义(t=4.983,P=0.008).抑制Wnt通路后,与阴性对照组相比,干扰组细胞凋亡率增加,对药物敏感性增强(P＜0.05).下调Wnt通路活性后p-ERK1/2的表达水平较阴性对照组明显降低,其他蛋白表达无明显差异.结论 抑制Wnt通路活性可部分逆转非小细胞肺癌细胞对吉非替尼的耐药性.
目的 探討下調Wnt通路活性對吉非替尼抑製非小細胞肺癌細胞株增殖的影響及其潛在的作用機製.方法 不同濃度吉非替尼處理PC9細胞株(吉非替尼敏感株)與PC9/AB2細胞株(吉非替尼耐藥株),細胞計數檢測試劑盒(CCK8)檢測兩種細胞株的增殖情況.Western blot分彆檢測PC9與PC9/AB2細胞中Wnt信號通路相關蛋白的錶達情況.雙熒光素酶報告基因繫統(TOP Flash)分彆檢測PC9和PC9/AB2細胞株Wnt通路的轉錄活性.採用β-catenin序列特異的siRNA抑製PC9/AB2中Wnt通路的轉錄活性後,用不同濃度吉非替尼處理,CCK8檢測榦預後細胞增殖情況.榦擾後錶皮生長因子受體及其下遊通路的錶達情況.結果 不同濃度吉非替尼作用下,PC9/AB2細胞對吉非替尼的耐藥性明顯增高(P＜0.05).PC9/AB2細胞Wnt通路相關蛋白的錶達明顯高于PC9,差異有統計學意義(t=24.590,P=0.000).TOP Flash結果顯示,PC9/AB2細胞株中Wnt通路的轉錄活性高于PC9細胞株,差異有統計學意義(t=4.983,P=0.008).抑製Wnt通路後,與陰性對照組相比,榦擾組細胞凋亡率增加,對藥物敏感性增彊(P＜0.05).下調Wnt通路活性後p-ERK1/2的錶達水平較陰性對照組明顯降低,其他蛋白錶達無明顯差異.結論 抑製Wnt通路活性可部分逆轉非小細胞肺癌細胞對吉非替尼的耐藥性.
목적 탐토하조Wnt통로활성대길비체니억제비소세포폐암세포주증식적영향급기잠재적작용궤제.방법 불동농도길비체니처리PC9세포주(길비체니민감주)여PC9/AB2세포주(길비체니내약주),세포계수검측시제합(CCK8)검측량충세포주적증식정황.Western blot분별검측PC9여PC9/AB2세포중Wnt신호통로상관단백적표체정황.쌍형광소매보고기인계통(TOP Flash)분별검측PC9화PC9/AB2세포주Wnt통로적전록활성.채용β-catenin서렬특이적siRNA억제PC9/AB2중Wnt통로적전록활성후,용불동농도길비체니처리,CCK8검측간예후세포증식정황.간우후표피생장인자수체급기하유통로적표체정황.결과 불동농도길비체니작용하,PC9/AB2세포대길비체니적내약성명현증고(P＜0.05).PC9/AB2세포Wnt통로상관단백적표체명현고우PC9,차이유통계학의의(t=24.590,P=0.000).TOP Flash결과현시,PC9/AB2세포주중Wnt통로적전록활성고우PC9세포주,차이유통계학의의(t=4.983,P=0.008).억제Wnt통로후,여음성대조조상비,간우조세포조망솔증가,대약물민감성증강(P＜0.05).하조Wnt통로활성후p-ERK1/2적표체수평교음성대조조명현강저,기타단백표체무명현차이.결론 억제Wnt통로활성가부분역전비소세포폐암세포대길비체니적내약성.
Objective To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib,and its related mechanisms.Methods PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib,and the proliferation index was measured using CCK8 kit.The members of Wnt signaling pathway were detected by Western blot.Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin.β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription,followed by treatment with different concentrations of gefitinib.Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.Results Treating with different concentrations of gefitinib,the resistance of PC9/AB2 cells to gefitinib was significantly increased (P ＜ 0.05).The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t =24.590,P =0.000).TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t =4.983,P =0.008).Compared with the negative control group,apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P ＜ 0.05).The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression,although other proteins showed no significant alterations.Conclusion Suppressing the activity of Wnt signaling can partly reverse the celluar risistance to gefitinib in non small cell lung cancer.
