中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2015年
6期
413-418
,共6页
朱丽华%徐刚%龚爱华%王燕%薛辉%张丽%吴朝阳
硃麗華%徐剛%龔愛華%王燕%薛輝%張麗%吳朝暘
주려화%서강%공애화%왕연%설휘%장려%오조양
Stattic%放射敏感性%肝癌细胞%细胞侵袭%细胞凋亡
Stattic%放射敏感性%肝癌細胞%細胞侵襲%細胞凋亡
Stattic%방사민감성%간암세포%세포침습%세포조망
Stattic%Radiosensitivity%Hepatocellular carcinoma cell%Cell invasion%Cell apoptosis
目的 观察信号转导子和转录激活子3(signal transducer and activator of transcription 3,STAT3)抑制剂Stattic(Y705)对人肝癌细胞系Bel-7402的生长、迁移、侵袭和放射敏感性的影响.方法 将人肝癌细胞系Bel-7402分为4组,空白对照组、Stattic组、单纯照射组和Stattic联合照射组(联合组).CCK8检测细胞的生长和增殖;划痕愈合实验和Transwell侵袭实验分别检测各组细胞的迁移能力和侵袭能力;克隆形成实验观察Stattic对Bel-7402细胞放射敏感性的影响;Western blot检测各组STAT3、p-STAT3、Bax,Bcl-2、Caspase-3、Cleaved Caspase-3、MMP-2、MMP-9蛋白的表达水平.结果 Stattic明显抑制人肝癌Bel-7402细胞的生长,并表现出剂量-效应关系,Stattic作用于细胞48 h后的IC50为2.5 μmol/L.1.0 μmol/L的Stattic与8 Gy射线照射联用,两者在细胞增殖抑制率上,表现出协同作用,抑制率下降了(15.00±1.87)%(F=63.30,P<0.05).划痕愈合实验显示联合组细胞的迁移能力明显降低,Transwell侵袭实验显示联合组滤膜细胞数明显减少.克隆形成实验显示,联合组与单纯照射组相比,克隆形成能力下降,SF2、D0、Dq均下降,(=4.20、6.92、9.32,P<0.05).Stattic在0.5 μmol/L时放射增敏比(SERSF2)为1.22.Western blot结果显示与单纯照射组相比,联合组的p-STAT3、MMP-2、MMP-9蛋白表达降低(=5.32、6.02、13.26,P<0.05).Bax、Cleaved Caspase-3蛋白表达升高(t=-7.82、-14.09,P<0.05).Bcl-2蛋白的表达下降(t=18.43,P <0.05).结论 Stattic通过抑制肝癌细胞Bel-7402的p-STAT3激活,一方面诱导凋亡增加肿瘤细胞的放射敏感性;另一方面降低了金属基质蛋白酶的上升,从而降低了肿瘤细胞迁移和侵袭能力.
目的 觀察信號轉導子和轉錄激活子3(signal transducer and activator of transcription 3,STAT3)抑製劑Stattic(Y705)對人肝癌細胞繫Bel-7402的生長、遷移、侵襲和放射敏感性的影響.方法 將人肝癌細胞繫Bel-7402分為4組,空白對照組、Stattic組、單純照射組和Stattic聯閤照射組(聯閤組).CCK8檢測細胞的生長和增殖;劃痕愈閤實驗和Transwell侵襲實驗分彆檢測各組細胞的遷移能力和侵襲能力;剋隆形成實驗觀察Stattic對Bel-7402細胞放射敏感性的影響;Western blot檢測各組STAT3、p-STAT3、Bax,Bcl-2、Caspase-3、Cleaved Caspase-3、MMP-2、MMP-9蛋白的錶達水平.結果 Stattic明顯抑製人肝癌Bel-7402細胞的生長,併錶現齣劑量-效應關繫,Stattic作用于細胞48 h後的IC50為2.5 μmol/L.1.0 μmol/L的Stattic與8 Gy射線照射聯用,兩者在細胞增殖抑製率上,錶現齣協同作用,抑製率下降瞭(15.00±1.87)%(F=63.30,P<0.05).劃痕愈閤實驗顯示聯閤組細胞的遷移能力明顯降低,Transwell侵襲實驗顯示聯閤組濾膜細胞數明顯減少.剋隆形成實驗顯示,聯閤組與單純照射組相比,剋隆形成能力下降,SF2、D0、Dq均下降,(=4.20、6.92、9.32,P<0.05).Stattic在0.5 μmol/L時放射增敏比(SERSF2)為1.22.Western blot結果顯示與單純照射組相比,聯閤組的p-STAT3、MMP-2、MMP-9蛋白錶達降低(=5.32、6.02、13.26,P<0.05).Bax、Cleaved Caspase-3蛋白錶達升高(t=-7.82、-14.09,P<0.05).Bcl-2蛋白的錶達下降(t=18.43,P <0.05).結論 Stattic通過抑製肝癌細胞Bel-7402的p-STAT3激活,一方麵誘導凋亡增加腫瘤細胞的放射敏感性;另一方麵降低瞭金屬基質蛋白酶的上升,從而降低瞭腫瘤細胞遷移和侵襲能力.
목적 관찰신호전도자화전록격활자3(signal transducer and activator of transcription 3,STAT3)억제제Stattic(Y705)대인간암세포계Bel-7402적생장、천이、침습화방사민감성적영향.방법 장인간암세포계Bel-7402분위4조,공백대조조、Stattic조、단순조사조화Stattic연합조사조(연합조).CCK8검측세포적생장화증식;화흔유합실험화Transwell침습실험분별검측각조세포적천이능력화침습능력;극륭형성실험관찰Stattic대Bel-7402세포방사민감성적영향;Western blot검측각조STAT3、p-STAT3、Bax,Bcl-2、Caspase-3、Cleaved Caspase-3、MMP-2、MMP-9단백적표체수평.결과 Stattic명현억제인간암Bel-7402세포적생장,병표현출제량-효응관계,Stattic작용우세포48 h후적IC50위2.5 μmol/L.1.0 μmol/L적Stattic여8 Gy사선조사련용,량자재세포증식억제솔상,표현출협동작용,억제솔하강료(15.00±1.87)%(F=63.30,P<0.05).화흔유합실험현시연합조세포적천이능력명현강저,Transwell침습실험현시연합조려막세포수명현감소.극륭형성실험현시,연합조여단순조사조상비,극륭형성능력하강,SF2、D0、Dq균하강,(=4.20、6.92、9.32,P<0.05).Stattic재0.5 μmol/L시방사증민비(SERSF2)위1.22.Western blot결과현시여단순조사조상비,연합조적p-STAT3、MMP-2、MMP-9단백표체강저(=5.32、6.02、13.26,P<0.05).Bax、Cleaved Caspase-3단백표체승고(t=-7.82、-14.09,P<0.05).Bcl-2단백적표체하강(t=18.43,P <0.05).결론 Stattic통과억제간암세포Bel-7402적p-STAT3격활,일방면유도조망증가종류세포적방사민감성;령일방면강저료금속기질단백매적상승,종이강저료종류세포천이화침습능력.
Objective To study the effects and preliminary mechanism of Stattic (Y705),an inhibitor of the signal transducer and activator of transcription-3 (STAT3),on the growth,migration,invasion and radiosensitization of hepatocellular carcinoma cell line Bel-7402.Methods Bel-7402 cells were divided to four groups:blank control group,Stattic treatment group,radiation group,and Stattic combined with radiation group.The cell growth and proliferation were detected by using CCK8 kit.The influence of Stattic on radiation sensitivity of Bel-7402 cells was determined by clone formation assay.The cell migration and invasion ability were tested by scratch migration assay and transwell assay,respectively.The protein expressions of STAT3,p-STAT3,Bax,Bcl-2,Caspase-3,Cleaved Caspase-3,MMP-2 and MMP-9 were quantified by Western blotting assay.Results Stattic significantly inhibited the growth of human hepatocellular carcinoma Bel-7402 cells with a dose-depended manner.The IC50 of Stattic after 48 h treatment was 2.5 μ mol/L.When 1.0 μmol/L Stattic was combined with 8 Gy X-rays,there was a synergistic effect in inhibition of cell proliferation with a inhibition rate of (15.00 ± 1.87) % (F =63.30,P < 0.05).Scratch migration assay and transwell invasion assay showed that the migration and invasion abilities of the combination group were significantly reduced.In addition,compared with the radiation group,the SF2,D0and Dq values obtained from survival curve were decreased (t =4.20,6.92,9.32,P <0.05),the protein expressions of p-STAT3,MMP-2,MMP-9 were reduced (t =5.32,6.02,13.26,P <0.05),the protein expressions of Bax and Cleaved Caspase-3 were increased in the combination group(t =-7.82,-14.09,P < 0.05),meanwhile the protein expressions of Bcl-2 was decreased (t =18.43,P < 0.05).When the concentration of Stattic was 0.5 μmol/L,the radiation sensitization ratio at 2 Gy (SERSF2) was 1.22.Conclusions By inhibiting the activation of the p-STAT3 in Bel-7402 cells,stattic could induce cell apoptosis and increase the radiosensitivity,down regulate MMP-2 and MMP-9 and thereby reduce the invasion and migration of tumor cells.