中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
4期
705-707
,共3页
颜畅%胡艺冰%穆磊%黄开禹%张哲%罗智勇
顏暢%鬍藝冰%穆磊%黃開禹%張哲%囉智勇
안창%호예빙%목뢰%황개우%장철%라지용
全反式维甲酸%结直肠癌%肿瘤干细胞%细胞分化
全反式維甲痠%結直腸癌%腫瘤榦細胞%細胞分化
전반식유갑산%결직장암%종류간세포%세포분화
All-trans retinoic acid%Colorectal cancer%Cancer stem cells%Cell differentiation
目的 探讨全反式维甲酸(ATRA)促进结直肠癌肿瘤干细胞分化的作用及其机制.方法 采用磁珠分选法从结直肠癌细胞株SW620细胞中分选出CD133+细胞,并用流式细胞仪检测1.0×10-6 mol/L ATRA处理3d后CD133+细胞比例的变化.采用成球实验观察1.0×10-6 mol/LATRA对结直肠癌干细胞处理后自我更新能力的影响.采用Western blot法检测1.0×10-6 mol/LATRA处理后β-链蛋白(β-catenin)及其磷酸化水平的改变.结果 SW620细胞中CD133+细胞富集肿瘤干细胞,CD133+和CD133-细胞的成球率分别为(51.00±7.67)％和(4.58±2.52)％(P＜0.01).ATRA能促进SW620细胞中CD133+细胞分化为CD133-细胞,用1.0×10-6mol/L ATRA处理细胞后,CD133+细胞的比例为(47.90 ±7.87)％,对照组CD133+细胞的比例为(95.33±2.24)％(P＜0.01).ATRA能抑制CD133+细胞的自我更新能力,ATRA处理组成球率为(12.67±2.08)％,对照组成球率为(24.33±2.52)％(P＜0.01).ATRA处理后CD133+细胞β-catenin的41号苏氨酸/45号丝氨酸位点(Thr41/Ser45)的磷酸化增加,β-catenin表达降低.结论 ATRA能促进结直肠癌干细胞中β-catenin的Thr41/Ser45位点磷酸化,促进β-catenin降解,抑制Wnt/β-catenin通路,诱导其分化为非肿瘤干细胞并降低自我更新能力.
目的 探討全反式維甲痠(ATRA)促進結直腸癌腫瘤榦細胞分化的作用及其機製.方法 採用磁珠分選法從結直腸癌細胞株SW620細胞中分選齣CD133+細胞,併用流式細胞儀檢測1.0×10-6 mol/L ATRA處理3d後CD133+細胞比例的變化.採用成毬實驗觀察1.0×10-6 mol/LATRA對結直腸癌榦細胞處理後自我更新能力的影響.採用Western blot法檢測1.0×10-6 mol/LATRA處理後β-鏈蛋白(β-catenin)及其燐痠化水平的改變.結果 SW620細胞中CD133+細胞富集腫瘤榦細胞,CD133+和CD133-細胞的成毬率分彆為(51.00±7.67)％和(4.58±2.52)％(P＜0.01).ATRA能促進SW620細胞中CD133+細胞分化為CD133-細胞,用1.0×10-6mol/L ATRA處理細胞後,CD133+細胞的比例為(47.90 ±7.87)％,對照組CD133+細胞的比例為(95.33±2.24)％(P＜0.01).ATRA能抑製CD133+細胞的自我更新能力,ATRA處理組成毬率為(12.67±2.08)％,對照組成毬率為(24.33±2.52)％(P＜0.01).ATRA處理後CD133+細胞β-catenin的41號囌氨痠/45號絲氨痠位點(Thr41/Ser45)的燐痠化增加,β-catenin錶達降低.結論 ATRA能促進結直腸癌榦細胞中β-catenin的Thr41/Ser45位點燐痠化,促進β-catenin降解,抑製Wnt/β-catenin通路,誘導其分化為非腫瘤榦細胞併降低自我更新能力.
목적 탐토전반식유갑산(ATRA)촉진결직장암종류간세포분화적작용급기궤제.방법 채용자주분선법종결직장암세포주SW620세포중분선출CD133+세포,병용류식세포의검측1.0×10-6 mol/L ATRA처리3d후CD133+세포비례적변화.채용성구실험관찰1.0×10-6 mol/LATRA대결직장암간세포처리후자아경신능력적영향.채용Western blot법검측1.0×10-6 mol/LATRA처리후β-련단백(β-catenin)급기린산화수평적개변.결과 SW620세포중CD133+세포부집종류간세포,CD133+화CD133-세포적성구솔분별위(51.00±7.67)％화(4.58±2.52)％(P＜0.01).ATRA능촉진SW620세포중CD133+세포분화위CD133-세포,용1.0×10-6mol/L ATRA처리세포후,CD133+세포적비례위(47.90 ±7.87)％,대조조CD133+세포적비례위(95.33±2.24)％(P＜0.01).ATRA능억제CD133+세포적자아경신능력,ATRA처리조성구솔위(12.67±2.08)％,대조조성구솔위(24.33±2.52)％(P＜0.01).ATRA처리후CD133+세포β-catenin적41호소안산/45호사안산위점(Thr41/Ser45)적린산화증가,β-catenin표체강저.결론 ATRA능촉진결직장암간세포중β-catenin적Thr41/Ser45위점린산화,촉진β-catenin강해,억제Wnt/β-catenin통로,유도기분화위비종류간세포병강저자아경신능력.
Objective To detect the promoting effects of all-trans retinoic acid (ATRA) on differentiation of cancer stem cells (CSCs) of colorectal cancer and the mechanism.Methods In order to enrich CSCs,magnetic activated cell sorting (MACS) was used to purify CD133 + CSCs from SW620 cell line.The proportion of CD133 + cells in purified CSCs was measured after treatment with 1.0 × 10-6 moL/L ATRA or dimethylsurfoxide (DMSO) for 3 days by flow cytometry.The inhibitory effect of self-renewing ability of CSCs induced by 1.0 × 10-6 mol/L ATRA was evaluated by sphere formation assay.Phosphorylation and degradation of β-catenin induced by 1.0 × 10-6 mol/L ATRA was confirmed by Western blotting.Results CD133 + SW620 cells effectively enrich CSCs.The rate of tumor spheres of CD133 + and CD133-cells was (51.00 ± 7.67) ％ and (4.58 ± 2.52) ％,respectively (P ＜ 0.01).After treatment with 1.0 × 10-6 mol/L ATRA,the proportion of CD133 + cells was (47.90 ±7.87)％,and that in the control group was (95.33 ± 2.24) ％,respectively (P ＜ 0.01).Interestingly,the rate of tumor spheres was decreased from (53.66 ± 6.03) ％ to (19.33 ± 2.52) ％ (P ＜ 0.01).Western blotting indicated that ATRA induced degradation of β-catenin by Thr41/Ser45 phosphorylation.Conclusion ATRA induces differentiation of colorectal CSCs,and inhibits their self-renewing ability by suppression of Wnt/β-catenin pathway.
