Cancerous inhibitor of protein phosphatase 2A(CIP2A) is a human oncoprotein that is overexpressed in multiple kinds of cancers including non-small cell lung cancer(NSCLC). CIP2A plays an ’oncogenic nexus’ to partici...Cancerous inhibitor of protein phosphatase 2A(CIP2A) is a human oncoprotein that is overexpressed in multiple kinds of cancers including non-small cell lung cancer(NSCLC). CIP2A plays an ’oncogenic nexus’ to participate in the tumorigenesis and chemoresistance in several cancer types. AKT and m TORC1 overactivation are detected in NSCLC and many other cancers. Previous studies found that the CIP2A/AKT/m TOR pathway controls cell growth, apoptosis, autophagy process. Polyphyllin I(PPI) and polyphyllin VII(PPVII) are natural components extracted from Paris polyphylla that display anti-cancer properties. In the present study, we investigated whether PPI and PPVII can be used in the cisplatin(DDP)-resistant human NSCLC cell line A549/DDP. Results demonstrated that PPI and PPVII treatment significantly suppressed A549/DDP cell proliferation, migration, invasion and EMT, induced apoptosis and autophagy. Further examination of the mechanism revealed that the PPI and PPVII significantly upregulated the p53, induced caspase-dependent apoptosis and suppressed the CIP2A/AKT/m TOR pathway. The activation of autophagy was mediated through PPI and PPVII induced inhibition of m TOR. We propose that PPI and PPVII might be developed as candidate drugs for DDP-resistant NSCLC.展开更多
目的:探讨miR-3163调控癌胚抗原相关黏附分子6(CEACAM6)对顺铂耐药胃癌细胞AGS/DDP增殖和凋亡的影响。方法:RT-qPCR和Western blot检测AGS/DDP细胞及亲本细胞株AGS中miR-3163和CEACAM6表达。将AGS/DDP细胞分为DDP+miR-NC、DDP+miR-3163...目的:探讨miR-3163调控癌胚抗原相关黏附分子6(CEACAM6)对顺铂耐药胃癌细胞AGS/DDP增殖和凋亡的影响。方法:RT-qPCR和Western blot检测AGS/DDP细胞及亲本细胞株AGS中miR-3163和CEACAM6表达。将AGS/DDP细胞分为DDP+miR-NC、DDP+miR-3163、DDP+si-NC、DDP+si-CEACAM6、DDP+miR-3163+pcDNA和DDP+miR-3163+pcDNACEACAM6组。MTT、流式细胞术分别检测细胞增殖和凋亡。双荧光素酶报告实验和Western blot确定miR-3163与CEACAM6的相互作用。结果:与AGS细胞比较,AGS/DDP细胞miR-3163表达显著降低(0.58±0.06 vs 1.00±0.06),CEACAM6表达显著升高(0.61±0.06 vs 0.24±0.03,P<0.05)。与DDP+miR-NC组比较,DDP+miR-3163组AGS/DDP细胞增殖抑制率[(36.32±3.21)%vs(8.41±0.83)%]、凋亡率[(23.14±2.33)%vs(7.55±0.76)%]显著升高(P<0.05)。与DDP+si-NC组比较,DDP+si-CEACAM6组AGS/DDP细胞增殖抑制率[(31.29±3.18)%vs(7.65±0.77)%]、凋亡率[(19.74±1.83)%vs(6.22±0.63)%]显著升高(P<0.05)。与DDP+miR-3163+pcDNA组比较,DDP+miR-3163+pcDNA-CEACAM6组AGS/DDP细胞增殖抑制率[(18.64±1.58)%vs(39.87±3.77)%]、凋亡率[(10.59±1.04)%vs(24.18±2.43)%]显著降低(P<0.05)。miR-3163靶向负调控CEACAM6表达。结论:miR-3163靶向CEACAM6抑制顺铂耐药胃癌细胞增殖,诱导细胞凋亡,提高其对顺铂的敏感性。展开更多
Objective To investigate whether low-dose fractionated radiation(LDFRT) could enhance cisplatin sensitivity in drug-resistant human ovarian cancer cells SKOV3/DDP, and to further explore the underlying mechanism.Metho...Objective To investigate whether low-dose fractionated radiation(LDFRT) could enhance cisplatin sensitivity in drug-resistant human ovarian cancer cells SKOV3/DDP, and to further explore the underlying mechanism.Methods SKOV3/DDP ovarian cancer cells were divided into three groups as follows: control, LDFRT, and conventional-dose radiation groups. Cells from all three groups were treated with different concentrations of cisplatin(0, 1.25, 2.5, 5, 10, and 20 μg/m L) for 48 h. The proliferation inhibition rate was investigated using the cell counting kit 8(CCK8). The rate of apoptosis was determined by flow cytometry(FCM). Protein levels of AKT, P-AKT, GSK-3β, P-GSK-3β, P21, cyclin D1, and P27 were examined by Western blotting. Results As expected, LDFRT significantly reduced the half-maximal inhibitory concentration(IC50) of cisplatin and promoted apoptosis in SKOV3/DDP cells. Moreover, in the LDFRT group, protein levels of P-AKT, P-GSK-3β, and cyclin D1 were markedly decreased, those of P21 and P27 were greatly increased, and total AKT and GSK-3β levels showed no significant difference compared to those in both the control and conventional-dose radiation groups.Conclusion LDFRT sensitizes resistant SKOV3/DDP ovarian cancer cells to cisplatin through inactivation of PI3 K/AKT/GSK-3β signaling.展开更多
目的分析MEK/ERK信号通路在上皮性卵巢癌顺铂耐药中的参与作用,为临床治疗提供参考。方法分析郑州大学第一附属医院2015年2月—2017年8月期间诊治的92例卵巢癌顺铂耐药患者(观察组)的临床资料,检测患者MEK/ERK(extracellular regulated ...目的分析MEK/ERK信号通路在上皮性卵巢癌顺铂耐药中的参与作用,为临床治疗提供参考。方法分析郑州大学第一附属医院2015年2月—2017年8月期间诊治的92例卵巢癌顺铂耐药患者(观察组)的临床资料,检测患者MEK/ERK(extracellular regulated protein kinases)信号转导通路相关蛋白的表达。以同期未发生顺铂耐药的卵巢癌患者(对照组)90例为对照。结果卵巢癌顺铂耐药患者血清中MEK/ERK信号通路蛋白Ras、Raf、MEK、ERK1、ERK2及PI3K和AKT蛋白的表达水平明显高于对照组,且差异存在统计学意义(均P<0.05);观察组患者血清中Bcl2(B-cell lymphoma-2)蛋白的表达水平明显高于对照组,细胞凋亡蛋白Bax(Bcl-2 associated X protein,BAX)和caspase-9(cysteinyl aspartate specific proteinase 9)水平明显低于对照组,且差异存在统计学意义(均P<0.05)。结论卵巢癌顺铂耐药过程可能与体内MEK/ERK信号通路及PI3K和AKT蛋白的异常表达有关。展开更多
To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the w...To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the wild PTEN gene expression plasmid (pGFP-PTEN) by magnetic iron nanoparticle and lipo2000. The transfection efficiency was detected by fluorescence microscope and flow cytometer. The expression levels of PTEN mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry analysis. The effect of PTEN transfection on cell cycle enhances the sensitivity of A549/CDDP to cisplatin and nanoparticle-mediated transfection has a higher efficiency than that of the liposome-mediated group. The apoptosis level was up-regulated in PTEN transfection group. The magnetic iron oxide nanoparticle could be used as one of the ideal gene carriers for PTEN gene delivery in vitro. PTEN can be an effective target for reversing cisplatin-resistance in lung cancer.展开更多
基金supported by the National Natural Science Foundation of China(No.81473485)the Natural Science Foundation of Shandong Province(No.2014ZRE27321)
文摘Cancerous inhibitor of protein phosphatase 2A(CIP2A) is a human oncoprotein that is overexpressed in multiple kinds of cancers including non-small cell lung cancer(NSCLC). CIP2A plays an ’oncogenic nexus’ to participate in the tumorigenesis and chemoresistance in several cancer types. AKT and m TORC1 overactivation are detected in NSCLC and many other cancers. Previous studies found that the CIP2A/AKT/m TOR pathway controls cell growth, apoptosis, autophagy process. Polyphyllin I(PPI) and polyphyllin VII(PPVII) are natural components extracted from Paris polyphylla that display anti-cancer properties. In the present study, we investigated whether PPI and PPVII can be used in the cisplatin(DDP)-resistant human NSCLC cell line A549/DDP. Results demonstrated that PPI and PPVII treatment significantly suppressed A549/DDP cell proliferation, migration, invasion and EMT, induced apoptosis and autophagy. Further examination of the mechanism revealed that the PPI and PPVII significantly upregulated the p53, induced caspase-dependent apoptosis and suppressed the CIP2A/AKT/m TOR pathway. The activation of autophagy was mediated through PPI and PPVII induced inhibition of m TOR. We propose that PPI and PPVII might be developed as candidate drugs for DDP-resistant NSCLC.
文摘目的:探讨miR-3163调控癌胚抗原相关黏附分子6(CEACAM6)对顺铂耐药胃癌细胞AGS/DDP增殖和凋亡的影响。方法:RT-qPCR和Western blot检测AGS/DDP细胞及亲本细胞株AGS中miR-3163和CEACAM6表达。将AGS/DDP细胞分为DDP+miR-NC、DDP+miR-3163、DDP+si-NC、DDP+si-CEACAM6、DDP+miR-3163+pcDNA和DDP+miR-3163+pcDNACEACAM6组。MTT、流式细胞术分别检测细胞增殖和凋亡。双荧光素酶报告实验和Western blot确定miR-3163与CEACAM6的相互作用。结果:与AGS细胞比较,AGS/DDP细胞miR-3163表达显著降低(0.58±0.06 vs 1.00±0.06),CEACAM6表达显著升高(0.61±0.06 vs 0.24±0.03,P<0.05)。与DDP+miR-NC组比较,DDP+miR-3163组AGS/DDP细胞增殖抑制率[(36.32±3.21)%vs(8.41±0.83)%]、凋亡率[(23.14±2.33)%vs(7.55±0.76)%]显著升高(P<0.05)。与DDP+si-NC组比较,DDP+si-CEACAM6组AGS/DDP细胞增殖抑制率[(31.29±3.18)%vs(7.65±0.77)%]、凋亡率[(19.74±1.83)%vs(6.22±0.63)%]显著升高(P<0.05)。与DDP+miR-3163+pcDNA组比较,DDP+miR-3163+pcDNA-CEACAM6组AGS/DDP细胞增殖抑制率[(18.64±1.58)%vs(39.87±3.77)%]、凋亡率[(10.59±1.04)%vs(24.18±2.43)%]显著降低(P<0.05)。miR-3163靶向负调控CEACAM6表达。结论:miR-3163靶向CEACAM6抑制顺铂耐药胃癌细胞增殖,诱导细胞凋亡,提高其对顺铂的敏感性。
文摘Objective To investigate whether low-dose fractionated radiation(LDFRT) could enhance cisplatin sensitivity in drug-resistant human ovarian cancer cells SKOV3/DDP, and to further explore the underlying mechanism.Methods SKOV3/DDP ovarian cancer cells were divided into three groups as follows: control, LDFRT, and conventional-dose radiation groups. Cells from all three groups were treated with different concentrations of cisplatin(0, 1.25, 2.5, 5, 10, and 20 μg/m L) for 48 h. The proliferation inhibition rate was investigated using the cell counting kit 8(CCK8). The rate of apoptosis was determined by flow cytometry(FCM). Protein levels of AKT, P-AKT, GSK-3β, P-GSK-3β, P21, cyclin D1, and P27 were examined by Western blotting. Results As expected, LDFRT significantly reduced the half-maximal inhibitory concentration(IC50) of cisplatin and promoted apoptosis in SKOV3/DDP cells. Moreover, in the LDFRT group, protein levels of P-AKT, P-GSK-3β, and cyclin D1 were markedly decreased, those of P21 and P27 were greatly increased, and total AKT and GSK-3β levels showed no significant difference compared to those in both the control and conventional-dose radiation groups.Conclusion LDFRT sensitizes resistant SKOV3/DDP ovarian cancer cells to cisplatin through inactivation of PI3 K/AKT/GSK-3β signaling.
文摘目的分析MEK/ERK信号通路在上皮性卵巢癌顺铂耐药中的参与作用,为临床治疗提供参考。方法分析郑州大学第一附属医院2015年2月—2017年8月期间诊治的92例卵巢癌顺铂耐药患者(观察组)的临床资料,检测患者MEK/ERK(extracellular regulated protein kinases)信号转导通路相关蛋白的表达。以同期未发生顺铂耐药的卵巢癌患者(对照组)90例为对照。结果卵巢癌顺铂耐药患者血清中MEK/ERK信号通路蛋白Ras、Raf、MEK、ERK1、ERK2及PI3K和AKT蛋白的表达水平明显高于对照组,且差异存在统计学意义(均P<0.05);观察组患者血清中Bcl2(B-cell lymphoma-2)蛋白的表达水平明显高于对照组,细胞凋亡蛋白Bax(Bcl-2 associated X protein,BAX)和caspase-9(cysteinyl aspartate specific proteinase 9)水平明显低于对照组,且差异存在统计学意义(均P<0.05)。结论卵巢癌顺铂耐药过程可能与体内MEK/ERK信号通路及PI3K和AKT蛋白的异常表达有关。
基金Project(07JJ3055)supported by the Natural Science Foundation of Hunan Province,China
文摘To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the wild PTEN gene expression plasmid (pGFP-PTEN) by magnetic iron nanoparticle and lipo2000. The transfection efficiency was detected by fluorescence microscope and flow cytometer. The expression levels of PTEN mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry analysis. The effect of PTEN transfection on cell cycle enhances the sensitivity of A549/CDDP to cisplatin and nanoparticle-mediated transfection has a higher efficiency than that of the liposome-mediated group. The apoptosis level was up-regulated in PTEN transfection group. The magnetic iron oxide nanoparticle could be used as one of the ideal gene carriers for PTEN gene delivery in vitro. PTEN can be an effective target for reversing cisplatin-resistance in lung cancer.
