目的探讨磷酸酶PHLPP1在人脐静脉内皮细胞(HUVEC)中的基础表达及转基因对其增殖的影响。方法体外培养的HUVEC分3组处理,分别为未转染组、转染pcDNA3-GFP组和转染pcDNA3HA-PHLPP组。通过构建pcDNA3HA-PHLPP1质粒并瞬时转染HUVEC。以细胞...目的探讨磷酸酶PHLPP1在人脐静脉内皮细胞(HUVEC)中的基础表达及转基因对其增殖的影响。方法体外培养的HUVEC分3组处理,分别为未转染组、转染pcDNA3-GFP组和转染pcDNA3HA-PHLPP组。通过构建pcDNA3HA-PHLPP1质粒并瞬时转染HUVEC。以细胞计数及噻唑盐比色法测定细胞增殖能力,Western blot ting定量磷酸酶PHLPP1蛋白表达水平。结果基础状态下HUVEC不表达PHLPP1。转染pcDNA3HA-PHLPP1组明显增加PHLPP1表达,与正常对照组、pcDNA3-GFP组比较差异显著(均P<0.01)。3组的细胞增殖指标无明显差异(P>0.05),其中MTT吸收度A值分别是0.134±0.015,0.133±0.014,0.137±0.016,细胞计数为(8.293±0.962)×105,(7.937±0.101)×105,8.127±0.112)×105。结论 PHLPP可能不是调节HUVEC增殖的最重要信号蛋白。展开更多
目的 :探讨PHLPP1及PHLPP2在食管上皮内瘤变及食管鳞癌组织中的表达及其临床意义。方法 :采用q RT-PCR方法检测40对食管上皮内瘤变及其正常对照组织和63对食管鳞癌及其正常对照组织中PHLPP1及PHLPP2 m RNA的表达情况,并探讨其表达量与...目的 :探讨PHLPP1及PHLPP2在食管上皮内瘤变及食管鳞癌组织中的表达及其临床意义。方法 :采用q RT-PCR方法检测40对食管上皮内瘤变及其正常对照组织和63对食管鳞癌及其正常对照组织中PHLPP1及PHLPP2 m RNA的表达情况,并探讨其表达量与临床病理资料的关系。结果:食管鳞癌组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于正常对照组织及上皮内瘤变组织(P<0.05)。食管上皮内瘤变组织中PHLPP1和PHLPP2 m RNA的表达水平与正常对照组织无统计学差异(P>0.05)。病理分级Ⅲ级的食管鳞癌患者组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于Ⅰ~Ⅱ级患者(P<0.01),TNM分期Ⅲ期的食管鳞癌患者组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于Ⅰ~Ⅱ期患者(P<0.01)。结论 :PHLPP1及PHLPP2表达水平与食管鳞癌发生发展与恶性程度密切相关,有望成为食管鳞癌诊断的新靶点。展开更多
Objective:Heat shock protein 47(HSP47)is a collagen-specific molecular chaperone that facilitates collagen maturation.Its role in cancer remains largely unknown.In this study,we investigated the roles o f HSP47 in col...Objective:Heat shock protein 47(HSP47)is a collagen-specific molecular chaperone that facilitates collagen maturation.Its role in cancer remains largely unknown.In this study,we investigated the roles o f HSP47 in colorectal cancer(CRC)and therapy resistance.Methods:Expression o f HSP47 in CRC tissues was examined(1)in paired human CRC/adjacent normal tissues,using real time quantitative reverse transcription polymerase chain reaction(qRT-PCR),The Cancer Genome Atlas(TCGA)database,and 22 independent m icroarray databases(curated CRC).In vitro studies on several CRC cell lines(H C T 116,RKO and C C L228)with modulated HSP47 expression were conducted to assess cell viability and apoptosis(TU N EL assay and caspase-3/-7)during exposure to chemotherapy.AKT signaling and co-imm unoprecipitation studies were performed to examine HSP47 and PHLPP1 interaction.In vivo studies using tumor xenografts were conducted to assess the effects of HSP47 modulation on tum or growth and therapy response.Results:HSP47 was upregulated in CRC and was associated with poor prognosis in individuals with CRC.In vitro,HSP47 overexpression supported the survival of CRC cells,whereas its knockdown sensitized cells to 5-fluorouracil(5-FU).HSP47 promoted survival by inhibiting apoptosis,enhancing AKT phosphorylation,and decreasing expression of the AKT-specific phosphatase PHLPP1 when cells were exposed to chemotherapy.These effects were partly results of the interaction between HSP47 and PHLPP1,which decreased PHLPP1 stability and led to more persistent AKT activity.In vivoy HSP47 supported tumor growth despite 5-FU treatment.Conclusions:HSP47 supports the growth of CRC tumors and suppresses the efficacy of chemotherapy via modulation of AKT signaling.展开更多
Objective To investigate the effects of insulin-like growth factor-1 （IGF-1） and oxidized low density lipoprotein （oxLDL） on expression ofphosphatase PHLPP 1 in vascular smooth muscle cells （VSMCs）. Methods Rabb...Objective To investigate the effects of insulin-like growth factor-1 （IGF-1） and oxidized low density lipoprotein （oxLDL） on expression ofphosphatase PHLPP 1 in vascular smooth muscle cells （VSMCs）. Methods Rabbit aortic VSMCs were cultured. VSMCs proliferation ability was determined by measuring cell number and mitochondrial dehydrogenase （MD） activity with MTT assay. Western blot was used to detect the protein expression ofphosphatase PHLPP1. Results IGF-1 （100ug/L） increased cell number and MD activity to 3.02 and 3.59 times of that in control group, oxLDL（501xg/ml） elevated the above two parameters to 2.03 and 2.91 times respectively. Western blot showed that IGF-1 and oxLDL inhibited the expression of PHLPPI to 39.27% and 40.26% of the control group （P〈0.01 ）. Conclusion IGF- 1 and oxLDL may enhance the proliferation of VSMCs by decreasing the expression ofphosphatase PHLPP 1.展开更多
文摘目的探讨磷酸酶PHLPP1在人脐静脉内皮细胞(HUVEC)中的基础表达及转基因对其增殖的影响。方法体外培养的HUVEC分3组处理,分别为未转染组、转染pcDNA3-GFP组和转染pcDNA3HA-PHLPP组。通过构建pcDNA3HA-PHLPP1质粒并瞬时转染HUVEC。以细胞计数及噻唑盐比色法测定细胞增殖能力,Western blot ting定量磷酸酶PHLPP1蛋白表达水平。结果基础状态下HUVEC不表达PHLPP1。转染pcDNA3HA-PHLPP1组明显增加PHLPP1表达,与正常对照组、pcDNA3-GFP组比较差异显著(均P<0.01)。3组的细胞增殖指标无明显差异(P>0.05),其中MTT吸收度A值分别是0.134±0.015,0.133±0.014,0.137±0.016,细胞计数为(8.293±0.962)×105,(7.937±0.101)×105,8.127±0.112)×105。结论 PHLPP可能不是调节HUVEC增殖的最重要信号蛋白。
文摘目的探讨PHLPP1在结直肠癌中的表达规律及临床意义。方法采用免疫组织化学实验检测10例结直肠癌组织石蜡标本中PHLPP1蛋白水平的表达情况;同时运用western blot实验和实时荧光定量PCR实验检测10对新鲜结直肠癌组织及配对的癌旁组织中PHLPP1蛋白和mRNA的表达情况;另外运用实时荧光定量PCR实验检测52例新鲜结直肠癌组织中PHLPP1 mRNA的表达水平,并分析PHLPP1的表达水平与临床病理参数之间的关系。结果 western blot实验、免疫组织化学实验及实时荧光定量PCR实验的结果显示PHLPP1在结直肠癌组织中的表达水平均明显低于配对的癌旁组织,并且PHLPP1在结直肠癌组织中mRNA的表达水平与结直肠癌的分化程度、Ducks分期、淋巴结转移和远处转移相关等临床病理参数之间呈负相关(P<0.05)。结论结直肠癌组织中PHLPP1的表达水平显著下调,并且其表达水平与分化程度、Dukes分期、淋巴结转移和远处转移等临床病理参数之间呈负相关,PHLPP1在结直肠癌的发生、发展和转移的过程中可能发挥类似抑癌基因的作用。
文摘目的 :探讨PHLPP1及PHLPP2在食管上皮内瘤变及食管鳞癌组织中的表达及其临床意义。方法 :采用q RT-PCR方法检测40对食管上皮内瘤变及其正常对照组织和63对食管鳞癌及其正常对照组织中PHLPP1及PHLPP2 m RNA的表达情况,并探讨其表达量与临床病理资料的关系。结果:食管鳞癌组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于正常对照组织及上皮内瘤变组织(P<0.05)。食管上皮内瘤变组织中PHLPP1和PHLPP2 m RNA的表达水平与正常对照组织无统计学差异(P>0.05)。病理分级Ⅲ级的食管鳞癌患者组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于Ⅰ~Ⅱ级患者(P<0.01),TNM分期Ⅲ期的食管鳞癌患者组织中PHLPP1和PHLPP2 m RNA的表达水平均明显低于Ⅰ~Ⅱ期患者(P<0.01)。结论 :PHLPP1及PHLPP2表达水平与食管鳞癌发生发展与恶性程度密切相关,有望成为食管鳞癌诊断的新靶点。
基金the Canadian Institutes of Health Research(CIHR)(Grant No.MOP-82881)the CIHR New Investigator Salary Award to Isabella T.Tai(Grant No.MSH-95344).
文摘Objective:Heat shock protein 47(HSP47)is a collagen-specific molecular chaperone that facilitates collagen maturation.Its role in cancer remains largely unknown.In this study,we investigated the roles o f HSP47 in colorectal cancer(CRC)and therapy resistance.Methods:Expression o f HSP47 in CRC tissues was examined(1)in paired human CRC/adjacent normal tissues,using real time quantitative reverse transcription polymerase chain reaction(qRT-PCR),The Cancer Genome Atlas(TCGA)database,and 22 independent m icroarray databases(curated CRC).In vitro studies on several CRC cell lines(H C T 116,RKO and C C L228)with modulated HSP47 expression were conducted to assess cell viability and apoptosis(TU N EL assay and caspase-3/-7)during exposure to chemotherapy.AKT signaling and co-imm unoprecipitation studies were performed to examine HSP47 and PHLPP1 interaction.In vivo studies using tumor xenografts were conducted to assess the effects of HSP47 modulation on tum or growth and therapy response.Results:HSP47 was upregulated in CRC and was associated with poor prognosis in individuals with CRC.In vitro,HSP47 overexpression supported the survival of CRC cells,whereas its knockdown sensitized cells to 5-fluorouracil(5-FU).HSP47 promoted survival by inhibiting apoptosis,enhancing AKT phosphorylation,and decreasing expression of the AKT-specific phosphatase PHLPP1 when cells were exposed to chemotherapy.These effects were partly results of the interaction between HSP47 and PHLPP1,which decreased PHLPP1 stability and led to more persistent AKT activity.In vivoy HSP47 supported tumor growth despite 5-FU treatment.Conclusions:HSP47 supports the growth of CRC tumors and suppresses the efficacy of chemotherapy via modulation of AKT signaling.
文摘Objective To investigate the effects of insulin-like growth factor-1 （IGF-1） and oxidized low density lipoprotein （oxLDL） on expression ofphosphatase PHLPP 1 in vascular smooth muscle cells （VSMCs）. Methods Rabbit aortic VSMCs were cultured. VSMCs proliferation ability was determined by measuring cell number and mitochondrial dehydrogenase （MD） activity with MTT assay. Western blot was used to detect the protein expression ofphosphatase PHLPP1. Results IGF-1 （100ug/L） increased cell number and MD activity to 3.02 and 3.59 times of that in control group, oxLDL（501xg/ml） elevated the above two parameters to 2.03 and 2.91 times respectively. Western blot showed that IGF-1 and oxLDL inhibited the expression of PHLPPI to 39.27% and 40.26% of the control group （P〈0.01 ）. Conclusion IGF- 1 and oxLDL may enhance the proliferation of VSMCs by decreasing the expression ofphosphatase PHLPP 1.
