目的观察消痰通腑方对结肠癌SW480细胞增殖、迁移的影响,探讨其分子作用机制。方法采用不同浓度消痰通腑方作用结肠癌SW480细胞,CCK8法和平板克隆形成实验检测细胞增殖,Transwell小室实验检测细胞迁移能力,RT-q PCR检测磷脂酶A2(PLA2)...目的观察消痰通腑方对结肠癌SW480细胞增殖、迁移的影响,探讨其分子作用机制。方法采用不同浓度消痰通腑方作用结肠癌SW480细胞,CCK8法和平板克隆形成实验检测细胞增殖,Transwell小室实验检测细胞迁移能力,RT-q PCR检测磷脂酶A2(PLA2)和环氧合酶2(COX-2)m RNA表达,Western blot检测PLA2和COX2蛋白表达,RNA干扰技术沉默SW480细胞株中PLA2基因表达。结果消痰通腑方低、高剂量组较对照组结肠癌SW480细胞增殖能力明显减弱(P<0.05),克隆形成率及迁移能力明显下降(P<0.05,P<0.01),PLA2、COX-2 m RNA和蛋白表达均明显下调(P<0.05),并呈浓度依赖性;与质粒对照组和对照组比较,PLA2 sh RNA干扰组细胞PLA2和COX2蛋白表达明显下调(P<0.05)。结论消痰通腑方可抑制结肠癌SW480细胞增殖和迁移,其机制可能与PLA2及COX-2的表达下调有关。展开更多
目的探究羟基氯喹对人结肠癌SW480细胞增殖及凋亡的影响,并分析可能的机制。方法体外培养人结肠癌细胞系SW480细胞,随机分为空白组和低、中、高剂量实验组。空白组正常培养,低、中、高剂量实验组分别用加入含羟基氯喹终浓度为20,40,80μ...目的探究羟基氯喹对人结肠癌SW480细胞增殖及凋亡的影响,并分析可能的机制。方法体外培养人结肠癌细胞系SW480细胞,随机分为空白组和低、中、高剂量实验组。空白组正常培养,低、中、高剂量实验组分别用加入含羟基氯喹终浓度为20,40,80μg·mL^(-1)的细胞培养液。用噻唑蓝(MTT)法检测细胞增殖活性;用平板克隆实验检测细胞克隆形成率;用AnnexinV-FITC/PI双染法检测细胞凋亡情况;用蛋白质印迹法检测Toll样受体4(TLR4)、核转录因子kappa B p65(NF-κB p65)、半胱氨酸的天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平。结果空白组和低、中、高剂量实验组SW480细胞存活率分别为100%,(83.15±12.45)%,(62.01±9.32)%和(43.25±6.45)%;这4组的克隆形成率分别为(85.16±12.77)%,(65.28±9.78)%,(51.06±7.07)%和(37.11±5.57)%;这4组的凋亡率分别为(9.12±1.37)%,(16.72±2.51)%,(25.89±3.89)%和(39.75±5.97)%;这4组的TLR4蛋白相对表达水平分别为1.15±0.18,0.90±0.13,0.65±0.09和0.29±0.05;这4组的NF-κB p65蛋白相对表达水平分别为1.27±0.20,0.96±0.15,0.69±0.11和0.47±0.08;这4组的Caspase-3蛋白相对表达水平分别为0.43±0.07,0.66±0.09,1.01±0.16和1.38±0.21。上述指标,低、中、高剂量组与空白组比较,差异均有统计学意义(均P<0.05)。结论羟基氯喹可能通过抑制TLR4/NF-κB通路活化抑制人结肠癌SW480细胞增殖并诱导细胞凋亡。展开更多
[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colo...[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colon cancer SW480 cells was detected by MTT colorimetry.The pro-apoptotic effect of OSNQ on human colon cancer SW480 cells was detected by Annexin V-FITC/PI double staining.The changes in expression of apoptosis-related proteins were detected by Western blot.[Results]The results of MTT assay showed that OSNQ had a significant cytotoxic effect on colon cancer SW480 cells.The results of Western blot showed that OSNQ induced the apoptosis in colon cancer SW480 cells through promoting the expression of pro-apoptotic caspase-3 and inhibiting the expression of apoptosis-inhibiting protein Bcl-2.[Conclusions] OSNQ has a significant cytotoxic effect on colon cancer SW480 cells,and it induces the apoptosis of colon cancer SW480 cells by AKT signaling pathway.展开更多
文摘目的观察消痰通腑方对结肠癌SW480细胞增殖、迁移的影响,探讨其分子作用机制。方法采用不同浓度消痰通腑方作用结肠癌SW480细胞,CCK8法和平板克隆形成实验检测细胞增殖,Transwell小室实验检测细胞迁移能力,RT-q PCR检测磷脂酶A2(PLA2)和环氧合酶2(COX-2)m RNA表达,Western blot检测PLA2和COX2蛋白表达,RNA干扰技术沉默SW480细胞株中PLA2基因表达。结果消痰通腑方低、高剂量组较对照组结肠癌SW480细胞增殖能力明显减弱(P<0.05),克隆形成率及迁移能力明显下降(P<0.05,P<0.01),PLA2、COX-2 m RNA和蛋白表达均明显下调(P<0.05),并呈浓度依赖性;与质粒对照组和对照组比较,PLA2 sh RNA干扰组细胞PLA2和COX2蛋白表达明显下调(P<0.05)。结论消痰通腑方可抑制结肠癌SW480细胞增殖和迁移,其机制可能与PLA2及COX-2的表达下调有关。
文摘目的探究羟基氯喹对人结肠癌SW480细胞增殖及凋亡的影响,并分析可能的机制。方法体外培养人结肠癌细胞系SW480细胞,随机分为空白组和低、中、高剂量实验组。空白组正常培养,低、中、高剂量实验组分别用加入含羟基氯喹终浓度为20,40,80μg·mL^(-1)的细胞培养液。用噻唑蓝(MTT)法检测细胞增殖活性;用平板克隆实验检测细胞克隆形成率;用AnnexinV-FITC/PI双染法检测细胞凋亡情况;用蛋白质印迹法检测Toll样受体4(TLR4)、核转录因子kappa B p65(NF-κB p65)、半胱氨酸的天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平。结果空白组和低、中、高剂量实验组SW480细胞存活率分别为100%,(83.15±12.45)%,(62.01±9.32)%和(43.25±6.45)%;这4组的克隆形成率分别为(85.16±12.77)%,(65.28±9.78)%,(51.06±7.07)%和(37.11±5.57)%;这4组的凋亡率分别为(9.12±1.37)%,(16.72±2.51)%,(25.89±3.89)%和(39.75±5.97)%;这4组的TLR4蛋白相对表达水平分别为1.15±0.18,0.90±0.13,0.65±0.09和0.29±0.05;这4组的NF-κB p65蛋白相对表达水平分别为1.27±0.20,0.96±0.15,0.69±0.11和0.47±0.08;这4组的Caspase-3蛋白相对表达水平分别为0.43±0.07,0.66±0.09,1.01±0.16和1.38±0.21。上述指标,低、中、高剂量组与空白组比较,差异均有统计学意义(均P<0.05)。结论羟基氯喹可能通过抑制TLR4/NF-κB通路活化抑制人结肠癌SW480细胞增殖并诱导细胞凋亡。
基金Supported by the Multigrain Production and Processing Characteristic Discipline Construction ProjectPostdoctoral Scientific Research Foundation of Heilongjiang Province of China(LBH-Q13132)
文摘[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colon cancer SW480 cells was detected by MTT colorimetry.The pro-apoptotic effect of OSNQ on human colon cancer SW480 cells was detected by Annexin V-FITC/PI double staining.The changes in expression of apoptosis-related proteins were detected by Western blot.[Results]The results of MTT assay showed that OSNQ had a significant cytotoxic effect on colon cancer SW480 cells.The results of Western blot showed that OSNQ induced the apoptosis in colon cancer SW480 cells through promoting the expression of pro-apoptotic caspase-3 and inhibiting the expression of apoptosis-inhibiting protein Bcl-2.[Conclusions] OSNQ has a significant cytotoxic effect on colon cancer SW480 cells,and it induces the apoptosis of colon cancer SW480 cells by AKT signaling pathway.
