AIM:To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy.METHODS:Three gastric cancer cell lines with distinct levels of differentiati...AIM:To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy.METHODS:Three gastric cancer cell lines with distinct levels of differentiation were treated with various doses of rAd-p53 alone,oxaliplatin(OXA) alone,or a combination of both.Cell growth was assessed with an 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and the expression levels of p53,Bax and Bcl-2 were determined by immunohistochemistry.The presence of apoptosis and the expression of caspase-3 were determined using flow cytometry.RESULTS:Treatment with rAd-p53 or OXA alone inhibited gastric cancer cell growth in a time-and dosedependent manner;moreover,significant synergistic effects were observed when these treatments were combined.Immunohistochemical analysis demonstrated that treatment with rAd-p53 alone,OXA alone or combined treatment led to decreased Bcl-2 expression and increased Bax expression in gastric cancer cells.Furthermore,flow cytometry showed that rAd-p53 alone,OXA alone or combination treatment induced apoptosis of gastric cancer cells,which was accompanied by increased expression of caspase-3.CONCLUSION:rAd-p53 enhances the sensitivity of gastric cancer cells to chemotherapy by promoting apoptosis.Thus,our results suggest that p53 gene therapy combined with chemotherapy represents a novel avenue for gastric cancer treatment.展开更多
目的探讨r Ad-p53基因对耐化疗药物MGC-803胃癌细胞的抑制作用。方法体外培养经紫杉醇处理的MGC-803细胞(MGC-803/PTX细胞),将其分为观察组和耐药对照组:观察组分别加入50、100、200、400、800、1600MOI r Ad-p53注射液,耐药对照组加入...目的探讨r Ad-p53基因对耐化疗药物MGC-803胃癌细胞的抑制作用。方法体外培养经紫杉醇处理的MGC-803细胞(MGC-803/PTX细胞),将其分为观察组和耐药对照组:观察组分别加入50、100、200、400、800、1600MOI r Ad-p53注射液,耐药对照组加入相应体积的培养液RPMI-1640;应用MTT法检测细胞增殖抑制情况,应用流式细胞仪检测观察期细胞凋亡率和细胞周期。采用"爬片法"收集细胞,将其分为r Ad-p53处理组,敏感对照组及耐药对照组,r Ad-p53处理组加入400 MOI r Ad-p53注射液,敏感对照组及耐药对照组均加入对应量的RPMI-1640培养基。3组分别采用免疫组化法在光学显微镜下观察细胞内多耐药基因表达蛋白(MDR1-Pgp蛋白)染色颗粒,及免疫印迹法检测MDR1-Pgp蛋白表达量(灰度值)。结果 MTT法检测结果显示,r Ad-p53对MGC-803/PTX细胞增殖的抑制作用显著,其抑制率随着时间的增加而逐渐上升;流式细胞术检测结果显示,100、200、400 MOI r Adp53处理组MGC-803/PTX细胞周期被阻滞在G2/M期,且其效果具有剂量依赖性;不同浓度的r Ad-p53处理后,MGC-803/PTX细胞凋亡率显著高于耐药对照组(P均<0.05)且具有剂量依赖性。免疫印迹法表明,r Ad-p53处理组MDR1-Pgp蛋白质灰度值显著高于敏感对照组,显著低于耐药对照组(P均<0.05);免疫组化法显示r Ad-p53处理组细胞核蓝染,可见少量阳性颗粒,而耐药对照组细胞内见大量深染颗粒,敏感对照组无阳性颗粒。结论 r Adp53对胃癌MGC-803/PTX细胞的抑制效果显著,因此可降低化疗治疗中的多药耐药性,为胃癌治疗提供了新的治疗途径。展开更多
Objective To explore the possible mechanism of rAd-p53 involved in glucose metabolism by observing the different effects of rAd-p53 on high-fat diet(HFD)fed with low-dose streptozotocin(STZ)treated(HFD/STZ)mice(β-cel...Objective To explore the possible mechanism of rAd-p53 involved in glucose metabolism by observing the different effects of rAd-p53 on high-fat diet(HFD)fed with low-dose streptozotocin(STZ)treated(HFD/STZ)mice(β-cell dysfunction model)and db/db mice(genetic insulin resistance model).Methods The established HFD/STZ and db/db diabetic mice were respectively展开更多
基金Supported by Xuzhou Science and Technology Development Fund,No. XM07C039
文摘AIM:To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy.METHODS:Three gastric cancer cell lines with distinct levels of differentiation were treated with various doses of rAd-p53 alone,oxaliplatin(OXA) alone,or a combination of both.Cell growth was assessed with an 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and the expression levels of p53,Bax and Bcl-2 were determined by immunohistochemistry.The presence of apoptosis and the expression of caspase-3 were determined using flow cytometry.RESULTS:Treatment with rAd-p53 or OXA alone inhibited gastric cancer cell growth in a time-and dosedependent manner;moreover,significant synergistic effects were observed when these treatments were combined.Immunohistochemical analysis demonstrated that treatment with rAd-p53 alone,OXA alone or combined treatment led to decreased Bcl-2 expression and increased Bax expression in gastric cancer cells.Furthermore,flow cytometry showed that rAd-p53 alone,OXA alone or combination treatment induced apoptosis of gastric cancer cells,which was accompanied by increased expression of caspase-3.CONCLUSION:rAd-p53 enhances the sensitivity of gastric cancer cells to chemotherapy by promoting apoptosis.Thus,our results suggest that p53 gene therapy combined with chemotherapy represents a novel avenue for gastric cancer treatment.
文摘目的探讨r Ad-p53基因对耐化疗药物MGC-803胃癌细胞的抑制作用。方法体外培养经紫杉醇处理的MGC-803细胞(MGC-803/PTX细胞),将其分为观察组和耐药对照组:观察组分别加入50、100、200、400、800、1600MOI r Ad-p53注射液,耐药对照组加入相应体积的培养液RPMI-1640;应用MTT法检测细胞增殖抑制情况,应用流式细胞仪检测观察期细胞凋亡率和细胞周期。采用"爬片法"收集细胞,将其分为r Ad-p53处理组,敏感对照组及耐药对照组,r Ad-p53处理组加入400 MOI r Ad-p53注射液,敏感对照组及耐药对照组均加入对应量的RPMI-1640培养基。3组分别采用免疫组化法在光学显微镜下观察细胞内多耐药基因表达蛋白(MDR1-Pgp蛋白)染色颗粒,及免疫印迹法检测MDR1-Pgp蛋白表达量(灰度值)。结果 MTT法检测结果显示,r Ad-p53对MGC-803/PTX细胞增殖的抑制作用显著,其抑制率随着时间的增加而逐渐上升;流式细胞术检测结果显示,100、200、400 MOI r Adp53处理组MGC-803/PTX细胞周期被阻滞在G2/M期,且其效果具有剂量依赖性;不同浓度的r Ad-p53处理后,MGC-803/PTX细胞凋亡率显著高于耐药对照组(P均<0.05)且具有剂量依赖性。免疫印迹法表明,r Ad-p53处理组MDR1-Pgp蛋白质灰度值显著高于敏感对照组,显著低于耐药对照组(P均<0.05);免疫组化法显示r Ad-p53处理组细胞核蓝染,可见少量阳性颗粒,而耐药对照组细胞内见大量深染颗粒,敏感对照组无阳性颗粒。结论 r Adp53对胃癌MGC-803/PTX细胞的抑制效果显著,因此可降低化疗治疗中的多药耐药性,为胃癌治疗提供了新的治疗途径。
文摘Objective To explore the possible mechanism of rAd-p53 involved in glucose metabolism by observing the different effects of rAd-p53 on high-fat diet(HFD)fed with low-dose streptozotocin(STZ)treated(HFD/STZ)mice(β-cell dysfunction model)and db/db mice(genetic insulin resistance model).Methods The established HFD/STZ and db/db diabetic mice were respectively
