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
384-387,416
,共5页
龙孝斌%郭仪%卢衡凭%邓治洲%刘辉%刘晓飞%罗吉辉%李汉贤
龍孝斌%郭儀%盧衡憑%鄧治洲%劉輝%劉曉飛%囉吉輝%李漢賢
룡효빈%곽의%로형빙%산치주%류휘%류효비%라길휘%리한현
乳腺癌干细胞%RNA干扰%HIF-2α%DLK1%移植瘤
乳腺癌榦細胞%RNA榦擾%HIF-2α%DLK1%移植瘤
유선암간세포%RNA간우%HIF-2α%DLK1%이식류
Breast cancer stem cells%RNA interference%HIF-2α%DLK1%Xenograft tumors
目的 探讨RNA干扰(RNA interference,RNAi)沉默低氧诱导因子-2α(hypoxia induciblefactor-2α,HIF-2αα)基因下调DLK1表达对乳腺癌干细胞(breast cancer stem cells,BCSCs)裸鼠移植瘤生长的影响.方法 建立BCSCs裸鼠移植瘤模型,原位注射RNAi慢病毒载体和空载体慢病毒载体至裸鼠左右侧背部移植瘤组织中,定期观察成瘤时间、瘤体体积及重量,进一步绘制生长曲线及计算抑瘤率.HE染色检测移植瘤组织病理,RT-PCR和Western blot法检测移植瘤组织HIF-2α、DLK1及CD44 mRNA和蛋白表达.结果 成功建立BCSCs裸鼠移植瘤模型.与空载体组相比,RNAi组移植瘤生长速率、瘤体体积和重量均明显降低(15.1％ vs 21.6％,5.80 cm3 vs 3.30 cm3,3.0 g vs 1.7 g,P＜0.05),抑瘤率为43.1％；HE染色显示RNAi组出现明显缺血坏死；RT-PCR结果提示RNAi组移植瘤组织中HIF-2α、DLK1及CD44蛋白表达均明显下调(0.59 ±0.02 vs 0.21 ±0.01,0.32 ±0.01 vs 0.08 ±0.01,0.76 ±0.03 vs0.04 ±0.01,P＜0.05)；Western blot结果也提示RNAi组移植瘤组织中HIF-2α、DLK1及CD44蛋白表达均明显下调(0.49 ±0.02 vs 0.28 ±0.01,0.43 ±0.02 vs0.11 ±0.01,0.62±0.03 vs 0.23±0.01,P＜0.05).结论 沉默HIF-2α基因下调DLK1表达有效抑制BCSCs裸鼠移植瘤生长,DLK1可能参与BCSCs未分化表型维持,并增强其致瘤性.
目的 探討RNA榦擾(RNA interference,RNAi)沉默低氧誘導因子-2α(hypoxia induciblefactor-2α,HIF-2αα)基因下調DLK1錶達對乳腺癌榦細胞(breast cancer stem cells,BCSCs)裸鼠移植瘤生長的影響.方法 建立BCSCs裸鼠移植瘤模型,原位註射RNAi慢病毒載體和空載體慢病毒載體至裸鼠左右側揹部移植瘤組織中,定期觀察成瘤時間、瘤體體積及重量,進一步繪製生長麯線及計算抑瘤率.HE染色檢測移植瘤組織病理,RT-PCR和Western blot法檢測移植瘤組織HIF-2α、DLK1及CD44 mRNA和蛋白錶達.結果 成功建立BCSCs裸鼠移植瘤模型.與空載體組相比,RNAi組移植瘤生長速率、瘤體體積和重量均明顯降低(15.1％ vs 21.6％,5.80 cm3 vs 3.30 cm3,3.0 g vs 1.7 g,P＜0.05),抑瘤率為43.1％；HE染色顯示RNAi組齣現明顯缺血壞死；RT-PCR結果提示RNAi組移植瘤組織中HIF-2α、DLK1及CD44蛋白錶達均明顯下調(0.59 ±0.02 vs 0.21 ±0.01,0.32 ±0.01 vs 0.08 ±0.01,0.76 ±0.03 vs0.04 ±0.01,P＜0.05)；Western blot結果也提示RNAi組移植瘤組織中HIF-2α、DLK1及CD44蛋白錶達均明顯下調(0.49 ±0.02 vs 0.28 ±0.01,0.43 ±0.02 vs0.11 ±0.01,0.62±0.03 vs 0.23±0.01,P＜0.05).結論 沉默HIF-2α基因下調DLK1錶達有效抑製BCSCs裸鼠移植瘤生長,DLK1可能參與BCSCs未分化錶型維持,併增彊其緻瘤性.
목적 탐토RNA간우(RNA interference,RNAi)침묵저양유도인자-2α(hypoxia induciblefactor-2α,HIF-2αα)기인하조DLK1표체대유선암간세포(breast cancer stem cells,BCSCs)라서이식류생장적영향.방법 건립BCSCs라서이식류모형,원위주사RNAi만병독재체화공재체만병독재체지라서좌우측배부이식류조직중,정기관찰성류시간、류체체적급중량,진일보회제생장곡선급계산억류솔.HE염색검측이식류조직병리,RT-PCR화Western blot법검측이식류조직HIF-2α、DLK1급CD44 mRNA화단백표체.결과 성공건립BCSCs라서이식류모형.여공재체조상비,RNAi조이식류생장속솔、류체체적화중량균명현강저(15.1％ vs 21.6％,5.80 cm3 vs 3.30 cm3,3.0 g vs 1.7 g,P＜0.05),억류솔위43.1％；HE염색현시RNAi조출현명현결혈배사；RT-PCR결과제시RNAi조이식류조직중HIF-2α、DLK1급CD44단백표체균명현하조(0.59 ±0.02 vs 0.21 ±0.01,0.32 ±0.01 vs 0.08 ±0.01,0.76 ±0.03 vs0.04 ±0.01,P＜0.05)；Western blot결과야제시RNAi조이식류조직중HIF-2α、DLK1급CD44단백표체균명현하조(0.49 ±0.02 vs 0.28 ±0.01,0.43 ±0.02 vs0.11 ±0.01,0.62±0.03 vs 0.23±0.01,P＜0.05).결론 침묵HIF-2α기인하조DLK1표체유효억제BCSCs라서이식류생장,DLK1가능삼여BCSCs미분화표형유지,병증강기치류성.
Objective To explore the effects of HIF-2α gene silence by small interference RNA on growth of breast cancer stem cells(BCSCs) in nude mice.Methods A nude mice xenograft tumors model with BCSCs was established.RNAi vector and empty vector lentiviral expression vector were subcutaneously injected into xenograft tumor tissues in the left and right flank of nude mice.The mean time of tumor growth and the volume and the weight of xenograft tumors were also measured at regular intervals.Furthermore,the growth curve was drawn and the inhibitory rate was calculated.The pathology of xenograft tumor tissues was detected by HE staining.mRNA and protein levels of HIF-2α,DLK1 and CD44 in xenograft tumors were detected by RT-PCR and Western blot respectively.Results The model of nude mice with BCSCs was constructed successfully.In contrast to empty vector group,the growth rate and the volumes and the weights of xenograft tumors obviously decreased in HIF-2α-siRNA group(15.1％ vs 21.6％,5.80cm3 vs 3..30cm3,3.0g vs 1.7g,P ＜0.05).The inhibitory rate of tumors was 43.1％.HE staining of tumor tissues indicated that ischemic necrosis was obvious in HIF-2oα-siRNA group.The expression of HIF-2oα,DLK1 and CD44 apparently reduced at mRNA (0.59 ± 0.02 vs 0.21 ±0.01,0.32 ±0.01 vs 0.08 ±0.01,0.76 ±0.03 vs 0.04 ±0.01,P ＜0.05),and protein levels(0.49 ± 0.02 vs 0.28 ± 0.01,0.43 ± 0.02 vs 0.11± 0.01,0.62 ± 0.03 vs 0.23 ± 0.01,P ＜ 0.05).Conclusion Silencing HIF-2α gene can inhibit the growth of BCSCs in nude mice.DLK1 may play an important role in maintenance of un-differentiation phenotype and tumorigenicity.
