AIM To investigate the anticancer effect of a recombinant adenovirus-mediated p53(r Ad-p53) combined with 5-fluorouracil(5-FU) in human colon cancer resistant to 5-FU in vivo and the mechanism of r Ad-p53 in reversal ...AIM To investigate the anticancer effect of a recombinant adenovirus-mediated p53(r Ad-p53) combined with 5-fluorouracil(5-FU) in human colon cancer resistant to 5-FU in vivo and the mechanism of r Ad-p53 in reversal of 5-FU resistance.METHODS nude mice bearing human colon cancer SW480/5-FU(5-FU resistant) were randomly assigned to four groups(n = 25 each): control group, 5-FU group, r Ad-p53 group, and r Ad-p53 + 5-FU group. At 24 h, 48 h, 72 h, 120 h and 168 h after treatment, 5 mice were randomly selected from each group and sacrificed using an overdose of anesthetics. The tumors were removed and the protein expressions of p53, protein kinase C(PKC), permeability-glycoprotein(P-gp) and multidrug resistance-associated protein 1(MRP1)(Western blot) and apoptosis(TUNEL) were determined.RESULTS The area ratios of tumor cell apoptosis were larger in the r Ad/p53 + 5-FU group than that in the control, 5-FU and r Ad/p53 groups(P < 0.05), and were larger in the r Ad/p53 group than that of the control group(P < 0.05) and the 5-FU group at more than 48 h(P < 0.05). The p53 expression was higher in the r Ad/p53 and the r Ad/p53 + 5-FU groups than that of the control and 5-FU groups(P < 0.05), and were higher in the r Ad/p53 + 5-FU group than that of the r Ad/p53 group(P < 0.05). Overexpression of PKC, P-gp and MRP1 was observed in the 5-FU and control groups. In the r Ad/p53 + 5-FU group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups(P < 0.05), and the expression of PKC was lower than that of the control, 5-FU and r Ad/p53 groups at more than 48 h(P < 0.05). In the r Ad/p53 group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups at more than 48 h(P < 0.05), and the expression of PKC was lower than that of the control and 5-FU groups at more than 120 h(P < 0.05).CONCLUSION5-FU combined with r Ad-p53 has a synergistic anticancer effect in SW480/5-FU(5-FU resistance), which contributes to reversal of 5-FU resistance.展开更多
目的探讨人结肠癌来源的间充质干细胞所分泌的外泌体对结肠癌细胞生物学功能的影响。方法将人结肠癌干细胞诱导培养出人结肠癌间充质干细胞(human colon cancer mesenchymal stem cells,hCC-MSCs),利用流式细胞学和成骨、成脂诱导分化...目的探讨人结肠癌来源的间充质干细胞所分泌的外泌体对结肠癌细胞生物学功能的影响。方法将人结肠癌干细胞诱导培养出人结肠癌间充质干细胞(human colon cancer mesenchymal stem cells,hCC-MSCs),利用流式细胞学和成骨、成脂诱导分化实验验证间充质干细胞的多能性。利用ExoQuick试剂盒提取hCC-MSCs外泌体(exosomes secreted from human colon cancer mesenchymal stem cells,hCC-MSCs-Exo),通过电镜、粒径分析和Western blot进行鉴定。采用PKH67标记hCC-MSCs-Exo,与HCT-116和HT-29共培养,荧光显微镜观察结肠癌细胞株摄取外泌体的情况,最后利用CCK8增殖实验、细胞划痕实验和Transwell侵袭实验检测hCC-MSCs-Exo对结肠癌细胞株生物学功能的影响。结果hCC-MSCs细胞呈单层贴壁生长,阳性表达CD29、CD90及CD105,阴性表达CD34、CD45及HLA-DR,具有成骨分化和成脂分化的多能性。hCC-MSCs-Exo平均直径135 nm,阳性表达Alix、CD63及CD81。hCC-MSCs-Exo与结肠癌细胞株共培养后,结肠癌细胞株能够摄取hCC-MSCs-Exo,且主要分布于细胞质中。CCK8、划痕实验和Transwell实验表明,与对照组相比,共培养组结肠癌细胞的增殖能力、迁移能力和侵袭能力显著增强(P<0.05)。结论hCC-MSCs-Exo能够促进结肠癌细胞增殖、迁徙和侵袭,为结肠癌药物治疗提供潜在的新治疗靶点。展开更多
基金Supported by the Natural Science Foundation of Guangdong,No.2015A030313732
文摘AIM To investigate the anticancer effect of a recombinant adenovirus-mediated p53(r Ad-p53) combined with 5-fluorouracil(5-FU) in human colon cancer resistant to 5-FU in vivo and the mechanism of r Ad-p53 in reversal of 5-FU resistance.METHODS nude mice bearing human colon cancer SW480/5-FU(5-FU resistant) were randomly assigned to four groups(n = 25 each): control group, 5-FU group, r Ad-p53 group, and r Ad-p53 + 5-FU group. At 24 h, 48 h, 72 h, 120 h and 168 h after treatment, 5 mice were randomly selected from each group and sacrificed using an overdose of anesthetics. The tumors were removed and the protein expressions of p53, protein kinase C(PKC), permeability-glycoprotein(P-gp) and multidrug resistance-associated protein 1(MRP1)(Western blot) and apoptosis(TUNEL) were determined.RESULTS The area ratios of tumor cell apoptosis were larger in the r Ad/p53 + 5-FU group than that in the control, 5-FU and r Ad/p53 groups(P < 0.05), and were larger in the r Ad/p53 group than that of the control group(P < 0.05) and the 5-FU group at more than 48 h(P < 0.05). The p53 expression was higher in the r Ad/p53 and the r Ad/p53 + 5-FU groups than that of the control and 5-FU groups(P < 0.05), and were higher in the r Ad/p53 + 5-FU group than that of the r Ad/p53 group(P < 0.05). Overexpression of PKC, P-gp and MRP1 was observed in the 5-FU and control groups. In the r Ad/p53 + 5-FU group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups(P < 0.05), and the expression of PKC was lower than that of the control, 5-FU and r Ad/p53 groups at more than 48 h(P < 0.05). In the r Ad/p53 group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups at more than 48 h(P < 0.05), and the expression of PKC was lower than that of the control and 5-FU groups at more than 120 h(P < 0.05).CONCLUSION5-FU combined with r Ad-p53 has a synergistic anticancer effect in SW480/5-FU(5-FU resistance), which contributes to reversal of 5-FU resistance.
文摘目的探讨人结肠癌来源的间充质干细胞所分泌的外泌体对结肠癌细胞生物学功能的影响。方法将人结肠癌干细胞诱导培养出人结肠癌间充质干细胞(human colon cancer mesenchymal stem cells,hCC-MSCs),利用流式细胞学和成骨、成脂诱导分化实验验证间充质干细胞的多能性。利用ExoQuick试剂盒提取hCC-MSCs外泌体(exosomes secreted from human colon cancer mesenchymal stem cells,hCC-MSCs-Exo),通过电镜、粒径分析和Western blot进行鉴定。采用PKH67标记hCC-MSCs-Exo,与HCT-116和HT-29共培养,荧光显微镜观察结肠癌细胞株摄取外泌体的情况,最后利用CCK8增殖实验、细胞划痕实验和Transwell侵袭实验检测hCC-MSCs-Exo对结肠癌细胞株生物学功能的影响。结果hCC-MSCs细胞呈单层贴壁生长,阳性表达CD29、CD90及CD105,阴性表达CD34、CD45及HLA-DR,具有成骨分化和成脂分化的多能性。hCC-MSCs-Exo平均直径135 nm,阳性表达Alix、CD63及CD81。hCC-MSCs-Exo与结肠癌细胞株共培养后,结肠癌细胞株能够摄取hCC-MSCs-Exo,且主要分布于细胞质中。CCK8、划痕实验和Transwell实验表明,与对照组相比,共培养组结肠癌细胞的增殖能力、迁移能力和侵袭能力显著增强(P<0.05)。结论hCC-MSCs-Exo能够促进结肠癌细胞增殖、迁徙和侵袭,为结肠癌药物治疗提供潜在的新治疗靶点。
