中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
2期
294-296
,共3页
肿瘤坏死因子诱导蛋白8样蛋白2%甲状腺癌%增殖%凋亡%侵袭
腫瘤壞死因子誘導蛋白8樣蛋白2%甲狀腺癌%增殖%凋亡%侵襲
종류배사인자유도단백8양단백2%갑상선암%증식%조망%침습
Tumor necrosis factor-α induced protein 8 like 2%Thyroid carcinoma cells%Proliferation%Apoptosis%Invasion
目的 检测肿瘤坏死因子诱导蛋白8样蛋白2(TIPE2)过表达对甲状腺癌细胞FTC-133增殖、凋亡以及侵袭能力的影响.方法 采用脂质体进行瞬时转染和G418筛选法构建TIPE2基因稳定转染甲状腺癌细胞FTC-133;细胞计数试剂盒(CCK-8)法检测24、48、72 h的细胞增殖,绘制细胞生长曲线,并计算细胞增殖抑制率;膜联蛋白V(Annexin V)/碘化丙锭(PI)法检测细胞凋亡,Transwell法检测细胞的侵袭能力.结果 筛选获得稳定高表达TIPE2的克隆细胞;Westernblot结果显示稳定转染TIPE2的FTC-133可明显上调TIPE2蛋白的表达水平,上调倍数达3.33倍;TIPE2过表达后细胞生长速度较对照组细胞明显减慢,24、48、72 h的细胞增殖抑制率分别为(21.60±5.66)％、(31.30±3.97)％、(40.80士7.37)％;TIPE2过表达后细胞凋亡明显增加,细胞总凋亡率为(13.06±1.89)％,而对照组细胞的总凋亡率为(5.73±0.43)％,两者差异有统计学意义(f=8.671,P ＜0.01);Transwell实验结果显示表达TIPE2的甲状腺癌细胞穿膜细胞数为(51.3 ±16.7)个,而对照组的穿膜细胞数为(91.7±22.7)个,差异有统计学意义(t=7.916,P＜0.01).结论 TIPE2能明显抑制甲状腺癌细胞的生长、促进其凋亡,同时抑制肿瘤细胞的侵袭能力.
目的 檢測腫瘤壞死因子誘導蛋白8樣蛋白2(TIPE2)過錶達對甲狀腺癌細胞FTC-133增殖、凋亡以及侵襲能力的影響.方法 採用脂質體進行瞬時轉染和G418篩選法構建TIPE2基因穩定轉染甲狀腺癌細胞FTC-133;細胞計數試劑盒(CCK-8)法檢測24、48、72 h的細胞增殖,繪製細胞生長麯線,併計算細胞增殖抑製率;膜聯蛋白V(Annexin V)/碘化丙錠(PI)法檢測細胞凋亡,Transwell法檢測細胞的侵襲能力.結果 篩選穫得穩定高錶達TIPE2的剋隆細胞;Westernblot結果顯示穩定轉染TIPE2的FTC-133可明顯上調TIPE2蛋白的錶達水平,上調倍數達3.33倍;TIPE2過錶達後細胞生長速度較對照組細胞明顯減慢,24、48、72 h的細胞增殖抑製率分彆為(21.60±5.66)％、(31.30±3.97)％、(40.80士7.37)％;TIPE2過錶達後細胞凋亡明顯增加,細胞總凋亡率為(13.06±1.89)％,而對照組細胞的總凋亡率為(5.73±0.43)％,兩者差異有統計學意義(f=8.671,P ＜0.01);Transwell實驗結果顯示錶達TIPE2的甲狀腺癌細胞穿膜細胞數為(51.3 ±16.7)箇,而對照組的穿膜細胞數為(91.7±22.7)箇,差異有統計學意義(t=7.916,P＜0.01).結論 TIPE2能明顯抑製甲狀腺癌細胞的生長、促進其凋亡,同時抑製腫瘤細胞的侵襲能力.
목적 검측종류배사인자유도단백8양단백2(TIPE2)과표체대갑상선암세포FTC-133증식、조망이급침습능력적영향.방법 채용지질체진행순시전염화G418사선법구건TIPE2기인은정전염갑상선암세포FTC-133;세포계수시제합(CCK-8)법검측24、48、72 h적세포증식,회제세포생장곡선,병계산세포증식억제솔;막련단백V(Annexin V)/전화병정(PI)법검측세포조망,Transwell법검측세포적침습능력.결과 사선획득은정고표체TIPE2적극륭세포;Westernblot결과현시은정전염TIPE2적FTC-133가명현상조TIPE2단백적표체수평,상조배수체3.33배;TIPE2과표체후세포생장속도교대조조세포명현감만,24、48、72 h적세포증식억제솔분별위(21.60±5.66)％、(31.30±3.97)％、(40.80사7.37)％;TIPE2과표체후세포조망명현증가,세포총조망솔위(13.06±1.89)％,이대조조세포적총조망솔위(5.73±0.43)％,량자차이유통계학의의(f=8.671,P ＜0.01);Transwell실험결과현시표체TIPE2적갑상선암세포천막세포수위(51.3 ±16.7)개,이대조조적천막세포수위(91.7±22.7)개,차이유통계학의의(t=7.916,P＜0.01).결론 TIPE2능명현억제갑상선암세포적생장、촉진기조망,동시억제종류세포적침습능력.
Objective To investigate the effect of tumor necrosis factor-α induced protein 8 like 2 (TIPE2) over-expression on cell proliferation,apoptosis and invasion in human thyroid carcinoma FTC-133 cells.Methods Stable transfected TIPE2 in FTC-133 cells was constructed using Lipofectamine 2000 transfection and G418 selection.Cell proliferation at 24,48 and 72 h was examined by cell counting kit-8 (CCK-8),cell growth curve and cell inhibitory rate were evaluated,cell apoptosis was examined by annexin V/propidium iodide (PI),and the cell invasion was evaluated using matrigel coated transwell assay.Results Western blotting analysis confirmed that TIPE2-stable transfected cells expressed high protein levels of TIPE2,which was 3.33 times higher than mock control cells.TIPE2 over expression inhibited cells growth with inhibitory rate being (21.60 ±5.66) ％,(31.30 ± 3.97)％、(40.80 ± 7.37) ％ at 24,48,72 h,respectively compared with mock control cells.TIPE2 over expression could promote cell apoptosis with apoptosis rate being (13.06 ± 1.89) ％,which was significantly higher than mock control cells [(5.73 ± 0.43) ％,t =8.671,P ＜ 0.01].The number of cells invading through matrigel was higher in the TIPE2 over-expressed cells (51.3 ± 16.7) than that in mock-transfected cells (91.7 ±22.7,t =7.916,P ＜ 0.01).Conclusion TIPE2 inhibited thyroid tumor growth,promoted tumor apoptosis and inhibited tumor cells invasion.
