中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2011年
6期
470-473
,共4页
肖朝文%彭志海%王从俊%于愿%陈堃%郑建伟%张俊%薛新波
肖朝文%彭誌海%王從俊%于願%陳堃%鄭建偉%張俊%薛新波
초조문%팽지해%왕종준%우원%진곤%정건위%장준%설신파
癌,肝细胞%溶瘤病毒%mda-7/IL-24基因%基因治疗
癌,肝細胞%溶瘤病毒%mda-7/IL-24基因%基因治療
암,간세포%용류병독%mda-7/IL-24기인%기인치료
Carcinoma,hepatocellular%Oncolytic viruses%Gene,mda-7/IL-24%Gene therapy
目的 观察携带人mda-7/IL-24的溶瘤腺病毒SG600-IL24对各种不同转移潜能肝癌细胞HepG2、SMMC7721、MHCC97L和正常肝细胞LO2的作用.方法将携带人mda-7/IL-24的溶瘤腺病毒SG600-IL24分别感染人肝癌细胞系HepG2、SMMC7721、MHCC97L和正常肝细胞LO2,逆转录-聚合酶链式反应(RT-PCR)、酶联免疫吸附试验(ELISA)、Western blot检测mda-7/IL-24基因和蛋白的表达,MTT观察肝癌细胞的生长抑制,Hoechst33258和流式细胞仪(FCM)检测细胞的凋亡,流式细胞仪(FCM)检测细胞周期.结果 RT-PCR和Western blot显示mda-7/IL-24在肝癌细胞和正常肝细胞中高表达,ELISA提示细胞培养上清液中mda-7/IL-24蛋白也明显增加.MTT和流式细胞仪提示mda-7/IL-24能明显抑制肝癌细胞的生长(生长抑制率分别为75%±2.5%,86%±3.5%,72%±1.8%,HepG2∶F=5.86,SMMC7721∶F=7.98,MHCC97L∶F=5.13,均P<0.01),促进肝癌细胞的凋亡(凋亡抑制率分别为56.5%±4.0%,34.4%±2.0%,43.3%±2.5%,HepG2∶F=203.4,SMMC7721∶F=130.5,MHCC97L∶F=160.6,均P<0.01),阻滞肝癌细胞在G2/M期(G2/M期阻滞分别为35.4%±4.2%,40.5%±5.0%,42.0%±5.0%,HepG2∶F=112.5,SMMC7721∶F=133.2,MHCC97L∶F=145.5,均P<0.01),而对正常肝细胞没有明显的促进凋亡和增殖阻滞作用.结论 溶瘤腺病毒SG600-IL24能特异性杀伤不同转移潜能人肝癌细胞和诱导凋亡,促进肝癌细胞增殖阻滞,而对正常肝细胞无明显促进凋亡和增殖阻滞作用.
目的 觀察攜帶人mda-7/IL-24的溶瘤腺病毒SG600-IL24對各種不同轉移潛能肝癌細胞HepG2、SMMC7721、MHCC97L和正常肝細胞LO2的作用.方法將攜帶人mda-7/IL-24的溶瘤腺病毒SG600-IL24分彆感染人肝癌細胞繫HepG2、SMMC7721、MHCC97L和正常肝細胞LO2,逆轉錄-聚閤酶鏈式反應(RT-PCR)、酶聯免疫吸附試驗(ELISA)、Western blot檢測mda-7/IL-24基因和蛋白的錶達,MTT觀察肝癌細胞的生長抑製,Hoechst33258和流式細胞儀(FCM)檢測細胞的凋亡,流式細胞儀(FCM)檢測細胞週期.結果 RT-PCR和Western blot顯示mda-7/IL-24在肝癌細胞和正常肝細胞中高錶達,ELISA提示細胞培養上清液中mda-7/IL-24蛋白也明顯增加.MTT和流式細胞儀提示mda-7/IL-24能明顯抑製肝癌細胞的生長(生長抑製率分彆為75%±2.5%,86%±3.5%,72%±1.8%,HepG2∶F=5.86,SMMC7721∶F=7.98,MHCC97L∶F=5.13,均P<0.01),促進肝癌細胞的凋亡(凋亡抑製率分彆為56.5%±4.0%,34.4%±2.0%,43.3%±2.5%,HepG2∶F=203.4,SMMC7721∶F=130.5,MHCC97L∶F=160.6,均P<0.01),阻滯肝癌細胞在G2/M期(G2/M期阻滯分彆為35.4%±4.2%,40.5%±5.0%,42.0%±5.0%,HepG2∶F=112.5,SMMC7721∶F=133.2,MHCC97L∶F=145.5,均P<0.01),而對正常肝細胞沒有明顯的促進凋亡和增殖阻滯作用.結論 溶瘤腺病毒SG600-IL24能特異性殺傷不同轉移潛能人肝癌細胞和誘導凋亡,促進肝癌細胞增殖阻滯,而對正常肝細胞無明顯促進凋亡和增殖阻滯作用.
목적 관찰휴대인mda-7/IL-24적용류선병독SG600-IL24대각충불동전이잠능간암세포HepG2、SMMC7721、MHCC97L화정상간세포LO2적작용.방법장휴대인mda-7/IL-24적용류선병독SG600-IL24분별감염인간암세포계HepG2、SMMC7721、MHCC97L화정상간세포LO2,역전록-취합매련식반응(RT-PCR)、매련면역흡부시험(ELISA)、Western blot검측mda-7/IL-24기인화단백적표체,MTT관찰간암세포적생장억제,Hoechst33258화류식세포의(FCM)검측세포적조망,류식세포의(FCM)검측세포주기.결과 RT-PCR화Western blot현시mda-7/IL-24재간암세포화정상간세포중고표체,ELISA제시세포배양상청액중mda-7/IL-24단백야명현증가.MTT화류식세포의제시mda-7/IL-24능명현억제간암세포적생장(생장억제솔분별위75%±2.5%,86%±3.5%,72%±1.8%,HepG2∶F=5.86,SMMC7721∶F=7.98,MHCC97L∶F=5.13,균P<0.01),촉진간암세포적조망(조망억제솔분별위56.5%±4.0%,34.4%±2.0%,43.3%±2.5%,HepG2∶F=203.4,SMMC7721∶F=130.5,MHCC97L∶F=160.6,균P<0.01),조체간암세포재G2/M기(G2/M기조체분별위35.4%±4.2%,40.5%±5.0%,42.0%±5.0%,HepG2∶F=112.5,SMMC7721∶F=133.2,MHCC97L∶F=145.5,균P<0.01),이대정상간세포몰유명현적촉진조망화증식조체작용.결론 용류선병독SG600-IL24능특이성살상불동전이잠능인간암세포화유도조망,촉진간암세포증식조체,이대정상간세포무명현촉진조망화증식조체작용.
Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24expressing human melanoma differentiation associated gene-7 (mda-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potential of HepG2, SMMC7721, MHCC97L and normal liver cell line LO2. Methods The oncolytic adenovirus SG600-IL24 which carrying mda-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line. The mRNA and protein expression of mda7/IL-24 in HepG2, SMMC7721, MHCC97L and LO2 cell lines was confirmed by RT-PCR,ELISA assay and Western blot respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst33258 and flow cytometry were studied to indicate the apoptosis effects. Results It was confirmed by RT-PCR, ELISA assay and Western-blot that the exogenous mda-7/IL-24 gene was highly expressed in HepG2, SMMC7721, MHCC97L and LO2 cell lines. MTT and apoptosis detection indicated that MDA-7/IL-24 can induce the growth suppression (the inhibition rate was 75% ±2. 5% ,86% ±3. 5% ,and promotes apoptosis ( the apoptosis rate was 56. 5% ± 4. 0% , 34. 4% ± 2. 0% , 43. 3% ± 2. 5%cell lines at G2/M phase ( the blocking rate was 35. 4% ± 4. 2% , 40. 5% ± 5. 0% , 42. 0% ± 5. 0%metastatic potential hepatocellular carcinoma cell lines but not in normal liver cell line.Conclusions Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression, promote apoptosis in hepatocellular carcinoma lines in vitro but not in normal liver cell LO2.
