中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
1期
18-20,封3
,共4页
周鹏成%王志伟%薄祥坤%朱慧%陆晶晶%李晓红%陆玉华
週鵬成%王誌偉%薄祥坤%硃慧%陸晶晶%李曉紅%陸玉華
주붕성%왕지위%박상곤%주혜%륙정정%리효홍%륙옥화
胰腺癌%肿瘤干细胞%八聚体结合转录因子4%胚胎干细胞转录因子%基因沉默
胰腺癌%腫瘤榦細胞%八聚體結閤轉錄因子4%胚胎榦細胞轉錄因子%基因沉默
이선암%종류간세포%팔취체결합전록인자4%배태간세포전록인자%기인침묵
Pancreatic cancer%Cancer stem cells%Octamer-binding transcription factor 4%Nanog%Gene silencing
目的 观察八聚体结合转录因子4(Oct4)和胚胎干细胞转录因子(Nanog)对胰腺癌干细胞(PCSCs)在体内干性特征的影响.方法 利用流式细胞分选技术分选人胰腺癌细胞-1(PANC-1)中CD44+ CD24+上皮特异性抗原(ESA)+胰腺癌干细胞,通过慢病毒载体介导特异性短发卡RNA (shRNA)沉默胰腺癌干细胞中的Oct4、Nanog基因,并通过Western blot检测干扰效率;将Oct4、Nanog沉默的胰腺癌干细胞、未沉默的胰腺癌干细胞及胰腺癌PANC-1细胞株分别接种于BALB/c裸鼠皮下及腹腔,建立裸鼠异位移植瘤模型,观察沉默Oct4、Nanog对胰腺癌干细胞体内致瘤性、侵袭性及耐药性的影响.结果 PANC-1细胞株中CD44+ CD24+ ESA+胰腺癌干细胞占细胞总数的0.1％ ～0.8％;shRNA沉默Oct4和Nanog的效率分别是(46.00 ±0.08)％和(78.00±0.12)％;体内实验结果显示,沉默Oct4、Nanog基因后,裸鼠的致瘤性和肿瘤转移性显著下降,对吉西他滨耐药性减弱.结论 沉默Oct4、Nanog表达可抑制裸鼠体内胰腺癌干细胞的干性特征.
目的 觀察八聚體結閤轉錄因子4(Oct4)和胚胎榦細胞轉錄因子(Nanog)對胰腺癌榦細胞(PCSCs)在體內榦性特徵的影響.方法 利用流式細胞分選技術分選人胰腺癌細胞-1(PANC-1)中CD44+ CD24+上皮特異性抗原(ESA)+胰腺癌榦細胞,通過慢病毒載體介導特異性短髮卡RNA (shRNA)沉默胰腺癌榦細胞中的Oct4、Nanog基因,併通過Western blot檢測榦擾效率;將Oct4、Nanog沉默的胰腺癌榦細胞、未沉默的胰腺癌榦細胞及胰腺癌PANC-1細胞株分彆接種于BALB/c裸鼠皮下及腹腔,建立裸鼠異位移植瘤模型,觀察沉默Oct4、Nanog對胰腺癌榦細胞體內緻瘤性、侵襲性及耐藥性的影響.結果 PANC-1細胞株中CD44+ CD24+ ESA+胰腺癌榦細胞佔細胞總數的0.1％ ～0.8％;shRNA沉默Oct4和Nanog的效率分彆是(46.00 ±0.08)％和(78.00±0.12)％;體內實驗結果顯示,沉默Oct4、Nanog基因後,裸鼠的緻瘤性和腫瘤轉移性顯著下降,對吉西他濱耐藥性減弱.結論 沉默Oct4、Nanog錶達可抑製裸鼠體內胰腺癌榦細胞的榦性特徵.
목적 관찰팔취체결합전록인자4(Oct4)화배태간세포전록인자(Nanog)대이선암간세포(PCSCs)재체내간성특정적영향.방법 이용류식세포분선기술분선인이선암세포-1(PANC-1)중CD44+ CD24+상피특이성항원(ESA)+이선암간세포,통과만병독재체개도특이성단발잡RNA (shRNA)침묵이선암간세포중적Oct4、Nanog기인,병통과Western blot검측간우효솔;장Oct4、Nanog침묵적이선암간세포、미침묵적이선암간세포급이선암PANC-1세포주분별접충우BALB/c라서피하급복강,건립라서이위이식류모형,관찰침묵Oct4、Nanog대이선암간세포체내치류성、침습성급내약성적영향.결과 PANC-1세포주중CD44+ CD24+ ESA+이선암간세포점세포총수적0.1％ ～0.8％;shRNA침묵Oct4화Nanog적효솔분별시(46.00 ±0.08)％화(78.00±0.12)％;체내실험결과현시,침묵Oct4、Nanog기인후,라서적치류성화종류전이성현저하강,대길서타빈내약성감약.결론 침묵Oct4、Nanog표체가억제라서체내이선암간세포적간성특정.
Objective To studythe impact of octamer-binding transcription factor 4 (Oct-4) and Nanog gene on the biological characteristics of pancreatic cancer stem cells (PCSCs) in vivo.Methods The CD44 + CD24 + epithelial specific antigen (ESA) + pancreatic cancer stem cells were sorted out via flow cytometry.The expression of Oct4 and Nanog genes in PCSCs were silenced by specific short hairpin RNA (shRNA) lentiviral vector.The interfering efficiency was detected by western blotting assay.The ectopic xenograft model was established via subcutaneous and intraperitoneal injection of Oct-4 and Nanog silenced PCSCs,normal PCSCs and PANC-1 cells into BALB/c nude mice,respectively.The influence of Oct4 and Nanog silencing on tumorigenicity,drug resistance and invasiveness were observed in vivo.Results The PCSCs accounted for 0.1％-0.8％ in PANC-1 cells.The efficiency of shRNA silencing Oct4 and Nanog were (46.00 ±0.08)％ and (78.00 ±0.12)％ respectively.Silencing of Oct4 and Nanog significantly inhibited the tumorigenicity and invasiveness of PCSCs in nude mice and reversed the resistance to gemcitabine.Conclusion The lentiviral vector-mediated silencing of Oct4 and Nanog could inhibit the stem cell-like biological characteristics of PCSCs in vivo.
