中华内分泌外科杂志
中華內分泌外科雜誌
중화내분비외과잡지
CHINESE JOURNAL OF ENDOCRINE SURGERY
2013年
5期
359-363
,共5页
杜成%刘兆喆%丁震宇%郭放%马东初%谢晓冬
杜成%劉兆喆%丁震宇%郭放%馬東初%謝曉鼕
두성%류조철%정진우%곽방%마동초%사효동
MTDH%小干扰RNA%乳腺癌%增殖%细胞周期%凋亡
MTDH%小榦擾RNA%乳腺癌%增殖%細胞週期%凋亡
MTDH%소간우RNA%유선암%증식%세포주기%조망
MTDH%siRNA%Breast cancer%Proliferation%Cell cycle%Apoptosis
目的 探讨转移粘附基因(metadherin,MTDH)表达下调对乳腺癌SK-BR-3细胞增殖和凋亡的影响及可能的机制.方法 采用RNA干扰技术,下调乳腺癌SK-BR-3细胞MTDH基因的表达,Western blot实验验证MTDH表达下调.四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium,MTT)实验检测MTDH下调对SK-BR-3细胞增殖的影响,流式细胞术检测细胞周期和凋亡情况.Western blot实验检测细胞增殖、周期和凋亡相关蛋白表达变化情况.结果 MTDH基因表达下调后,乳腺癌SK-BR-3细胞增殖能力减弱,作用48和72h后,空白对照组、阴性对照组和实验组吸光度A值分别为(2.0 ±0.1)vs(1.9±0.3) vs(0.9 ±0.1)(P =0.02)和(2.7±0.2) vs(2.5 ±0.4)vs(1.3±0.2)(P =0.008).细胞阻滞于G0/G1期,S期和G2/M期细胞比例下降.MTDH表达下调可诱导乳腺癌细胞凋亡,空白对照组、阴性对照组和实验组细胞的凋亡率分别为(1.3±0.2)％、(1.4±0.3)％和(19.6±2.7)％(P=0.012).MTDH表达下调可显著降低细胞cyclinD1和Bcl-2的表达,但P21表达升高,并可使caspase-3活化.结论 下调MTDH基因表达可抑制乳腺癌SK-BR-3细胞增殖并诱导其凋亡,其机制可能与cyclinD1和Bcl-2的表达下调、P21表达上调及caspase-3活化有关.
目的 探討轉移粘附基因(metadherin,MTDH)錶達下調對乳腺癌SK-BR-3細胞增殖和凋亡的影響及可能的機製.方法 採用RNA榦擾技術,下調乳腺癌SK-BR-3細胞MTDH基因的錶達,Western blot實驗驗證MTDH錶達下調.四甲基偶氮唑鹽比色法(methyl thiazolyl tetrazolium,MTT)實驗檢測MTDH下調對SK-BR-3細胞增殖的影響,流式細胞術檢測細胞週期和凋亡情況.Western blot實驗檢測細胞增殖、週期和凋亡相關蛋白錶達變化情況.結果 MTDH基因錶達下調後,乳腺癌SK-BR-3細胞增殖能力減弱,作用48和72h後,空白對照組、陰性對照組和實驗組吸光度A值分彆為(2.0 ±0.1)vs(1.9±0.3) vs(0.9 ±0.1)(P =0.02)和(2.7±0.2) vs(2.5 ±0.4)vs(1.3±0.2)(P =0.008).細胞阻滯于G0/G1期,S期和G2/M期細胞比例下降.MTDH錶達下調可誘導乳腺癌細胞凋亡,空白對照組、陰性對照組和實驗組細胞的凋亡率分彆為(1.3±0.2)％、(1.4±0.3)％和(19.6±2.7)％(P=0.012).MTDH錶達下調可顯著降低細胞cyclinD1和Bcl-2的錶達,但P21錶達升高,併可使caspase-3活化.結論 下調MTDH基因錶達可抑製乳腺癌SK-BR-3細胞增殖併誘導其凋亡,其機製可能與cyclinD1和Bcl-2的錶達下調、P21錶達上調及caspase-3活化有關.
목적 탐토전이점부기인(metadherin,MTDH)표체하조대유선암SK-BR-3세포증식화조망적영향급가능적궤제.방법 채용RNA간우기술,하조유선암SK-BR-3세포MTDH기인적표체,Western blot실험험증MTDH표체하조.사갑기우담서염비색법(methyl thiazolyl tetrazolium,MTT)실험검측MTDH하조대SK-BR-3세포증식적영향,류식세포술검측세포주기화조망정황.Western blot실험검측세포증식、주기화조망상관단백표체변화정황.결과 MTDH기인표체하조후,유선암SK-BR-3세포증식능력감약,작용48화72h후,공백대조조、음성대조조화실험조흡광도A치분별위(2.0 ±0.1)vs(1.9±0.3) vs(0.9 ±0.1)(P =0.02)화(2.7±0.2) vs(2.5 ±0.4)vs(1.3±0.2)(P =0.008).세포조체우G0/G1기,S기화G2/M기세포비례하강.MTDH표체하조가유도유선암세포조망,공백대조조、음성대조조화실험조세포적조망솔분별위(1.3±0.2)％、(1.4±0.3)％화(19.6±2.7)％(P=0.012).MTDH표체하조가현저강저세포cyclinD1화Bcl-2적표체,단P21표체승고,병가사caspase-3활화.결론 하조MTDH기인표체가억제유선암SK-BR-3세포증식병유도기조망,기궤제가능여cyclinD1화Bcl-2적표체하조、P21표체상조급caspase-3활화유관.
Objective To investigate if down regulation of MTDH could inhibit proliferation and induce apoptosis in breast cancer SK-BR-3 cells.Methods RNA interference was employed to reduce MTDH expression in human breast cancer SK-BR-3 cells.Western blot assay was applied to measure the down regulation of MTDH.MTT assay was performed to assess the proliferation of SK-BR-3 cell.Flow cytometry was employed to detect cell cycle and apoptosis.Western blot assay was applied to detect the molecular alterations that was associated with cell proliferation,cell cycle and apoptosis.Results MTDH down regulation significantly inhibited cell proliferation.48 hours and 72 hours after trasnfection,the absorbance value(A value)in blank control,negative control and treatment group was (2.0 ± 0.1) vs (1.9 ± 0.3) vs (0.9 ± 0.1) (P =0.02) and (2.7 ± 0.2) vs (2.5 ± 0.4) vs (1.3 ± 0.2) (P =0.008).MTDH down regulation resulted in accumulation of the G0/G1 phase cells and reduction of S and G2/M phase cells.Moreover,MTDH down regulation significantly induced cell apoptosis.The cell apoptosis rate in blank control,negative control and trial group was (1.3 ± 0.2) ％,(1.4 ± 0.3) ％ and (19.6 ± 2.7) ％ (P =0.012).MTDH down regulation resulted in a decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.Conclusions Reduced MTDH expression in SKBR-3 cells can inhibit proliferation and induce apoptosis,which may be associated with decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.
