中国医药
中國醫藥
중국의약
CHINA MEDICINE
2014年
10期
1492-1494
,共3页
肝癌%二甲双胍%表皮生长因子%细胞凋亡
肝癌%二甲雙胍%錶皮生長因子%細胞凋亡
간암%이갑쌍고%표피생장인자%세포조망
Liver cancer%Metformin%Epidermal growth factor%Apoptosis
目的 观察二甲双胍对人肝癌Hep-G2细胞表皮生长因子(EGF)及其受体(EGFR)的影响.方法 体外培养人肝癌Hep-G2细胞,用不同浓度二甲双胍进行干预,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氨唑溴盐法检测二甲双胍对Hep-G2细胞生长的影响;Hoechst 33342染色荧光显微镜观察细胞的形态学变化,流式细胞术观察细胞凋亡;蛋白质印迹法(Western blot)检测EGF、EGFR的表达.结果 二甲双胍对肝癌细胞的活性具有明显的抑制作用,且呈剂量依赖性;以10 mmol/L作用终浓度抑制效果最明显,其48 h的细胞吸光度为(0.477 ±0.025),与对照组(0.602±0.026)比较差异有统计学意义(P＜0.01);逐渐增加二甲双胍的作用浓度,细胞凋亡率随之逐渐增加,药物组细胞凋亡率分别为(10.76±0.96)％、(20.77±1.16)％,与对照组(5.21±0.13)％相比差异均有统计学意义(P＜0.05);Hoechst33342染色可见明显的细胞皱缩,核染色质浓缩,核碎裂等凋亡形态学变化;Western blot结果显示,EGF及其受体蛋白的表达随着二甲双胍作用浓度的增加而逐渐下调.结论 在体外,二甲双胍能够抑制人肝癌Hep-G2细胞增殖及诱导细胞凋亡,其抗肿瘤机制可能与细胞内EGF及EGFR表达下调有关.
目的 觀察二甲雙胍對人肝癌Hep-G2細胞錶皮生長因子(EGF)及其受體(EGFR)的影響.方法 體外培養人肝癌Hep-G2細胞,用不同濃度二甲雙胍進行榦預,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氨唑溴鹽法檢測二甲雙胍對Hep-G2細胞生長的影響;Hoechst 33342染色熒光顯微鏡觀察細胞的形態學變化,流式細胞術觀察細胞凋亡;蛋白質印跡法(Western blot)檢測EGF、EGFR的錶達.結果 二甲雙胍對肝癌細胞的活性具有明顯的抑製作用,且呈劑量依賴性;以10 mmol/L作用終濃度抑製效果最明顯,其48 h的細胞吸光度為(0.477 ±0.025),與對照組(0.602±0.026)比較差異有統計學意義(P＜0.01);逐漸增加二甲雙胍的作用濃度,細胞凋亡率隨之逐漸增加,藥物組細胞凋亡率分彆為(10.76±0.96)％、(20.77±1.16)％,與對照組(5.21±0.13)％相比差異均有統計學意義(P＜0.05);Hoechst33342染色可見明顯的細胞皺縮,覈染色質濃縮,覈碎裂等凋亡形態學變化;Western blot結果顯示,EGF及其受體蛋白的錶達隨著二甲雙胍作用濃度的增加而逐漸下調.結論 在體外,二甲雙胍能夠抑製人肝癌Hep-G2細胞增殖及誘導細胞凋亡,其抗腫瘤機製可能與細胞內EGF及EGFR錶達下調有關.
목적 관찰이갑쌍고대인간암Hep-G2세포표피생장인자(EGF)급기수체(EGFR)적영향.방법 체외배양인간암Hep-G2세포,용불동농도이갑쌍고진행간예,3-(4,5-이갑기새서-2)-2,5-이분기사안서추염법검측이갑쌍고대Hep-G2세포생장적영향;Hoechst 33342염색형광현미경관찰세포적형태학변화,류식세포술관찰세포조망;단백질인적법(Western blot)검측EGF、EGFR적표체.결과 이갑쌍고대간암세포적활성구유명현적억제작용,차정제량의뢰성;이10 mmol/L작용종농도억제효과최명현,기48 h적세포흡광도위(0.477 ±0.025),여대조조(0.602±0.026)비교차이유통계학의의(P＜0.01);축점증가이갑쌍고적작용농도,세포조망솔수지축점증가,약물조세포조망솔분별위(10.76±0.96)％、(20.77±1.16)％,여대조조(5.21±0.13)％상비차이균유통계학의의(P＜0.05);Hoechst33342염색가견명현적세포추축,핵염색질농축,핵쇄렬등조망형태학변화;Western blot결과현시,EGF급기수체단백적표체수착이갑쌍고작용농도적증가이축점하조.결론 재체외,이갑쌍고능구억제인간암Hep-G2세포증식급유도세포조망,기항종류궤제가능여세포내EGF급EGFR표체하조유관.
Objective To observe the effects of metformin on the expression of epidermal growth factor (EGF) and epidermal growth factor recipient (EGFR) in human liver cancer cell line Hep-G2.Methods In vitro,the Hep-G2 cells were treated with different concentrations of metformin.The proliferation of the cells was detected by MTT assay and the apoptosis of the cells was measured by cytochemical staining with Hoechst 33342 and flow cytometry.The expressions of EGF and EGFR were investigated by western blot.Results Metformin inhibited the growth of Hep-G2 cells obviously in a dose-dependent manner.The absorbance was (0.477 ± 0.025)after metformin was used for 48 h,which was significandy higher than that of the control patients (0.602 ± 0.026) (P ＜ 0.05).FCM analysis showed that when Hep-G2 cells were treated with metformin,the apoptosis rates were (10.76 ± 0.96) ％ and (20.77 ± 1.16) ％,respectively,compared with the control group (5.21 ± 0.13) ％ in creased (P ＜ 0.05).Cell apoptosis with cell shrinkage,nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed by cytochemical staining.The western-blot showed that the expressions of EGF and EGFR were down regulated while the concentration of metformin was increasing.Conclusion Metformin can inhibit cell proliferation and induce apoptosis in liver cancer cell line Hep-G2,which may be related to the down-regulation of EGF and EGFR protein expression.
