Objective:Physical exercise,a common non-drug intervention,is an important strategy in cancer treatment,including hepatocellular carcinoma(HCC).However,the mechanism remains largely unknown.Due to the importance of hy...Objective:Physical exercise,a common non-drug intervention,is an important strategy in cancer treatment,including hepatocellular carcinoma(HCC).However,the mechanism remains largely unknown.Due to the importance of hypoxia and cancer stemness in the development of HCC,the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness.Methods:A physical exercise intervention of swimming(30 min/d,5 d/week,for 4 weeks)was administered to BALB/c nude mice bearing subcutaneous human HCC tumor.The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring,hematoxylin and eosin(HE)staining,and immunohistochemistry(IHC)detection of proliferating cell nuclear antigen(PCNA)and Ki67.The expression of stemness transcription factors,including Nanog homeobox(NANOG),octamer-binding transcription factor 4(OCT-4),v-Myc avian myelocytomatosis viral oncogene homolog(C-MYC)and hypoxia-inducible factor-1a(HIF-1a),was detected using real-time reverse transcription polymerase chain reaction.A hypoxia probe was used to explore the intratumoral hypoxia status.Western blot was used to detect the expression of HIF-1a and proteins related to protein kinase B(Akt)/glycogen synthase kinase-3β(GSK-3β)/β-catenin signaling pathway.The IHC analysis of platelet endothelial cell adhesion molecule-1(CD31),and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion.SMMC-7721 cells were treated with nude mice serum.The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors.The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1a.Further,the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected.Results:Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor.HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention.Swimming potently alleviated the intratumoral hypoxia,attenuated the cancer stemness,and inhibited the Akt/GSK-3β/β-catenin signaling pathway.Additionally,the desmin+/CD31+ratio,rather than the number of CD31+vessels,was significantly increased in swimming-treated mice.In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere,as well as the m RNA expression level of stemness transcription factors.Consistent with the in vivo results,HIF-1a and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group.Conclusion:Swimming alleviated hypoxia and attenuated cancer stemness in HCC,through suppression of the Akt/GSK-3β/β-catenin signaling pathway.The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor.展开更多
目的探讨驱动蛋白家族成员11(kinesin family member 11,KIF11)、β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)在宫颈癌中的表达情况及临床意义。方法采用免疫组化法检测KIF11、β-catenin、GS...目的探讨驱动蛋白家族成员11(kinesin family member 11,KIF11)、β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)在宫颈癌中的表达情况及临床意义。方法采用免疫组化法检测KIF11、β-catenin、GSK-3β在102例宫颈癌、52例高级别鳞状上皮内病变(high-grade squamous intraepithelial lesion,HSIL)、46例低级别鳞状上皮内病变(low-grade squamous intraepithelial lesion,LSIL)及40例慢性宫颈炎组织中的表达情况,分析三者的表达与宫颈癌患者临床病理特征的关系,分析三者之间的相关性,COX比例风险模型分析影响宫颈癌患者预后的影响因素。结果随着宫颈病变进展,KIF11、β-catenin阳性率逐渐升高,GSK-3β阳性率逐渐减低(P<0.05)。KIF11、β-catenin、GSK-3β在宫颈癌组织中的阳性表达在国际妇产科联盟(International Federation of Gynecology and Obstetrics,FIGO)分期、分化程度、淋巴结转移方面比较,差异有统计学意义(P<0.05),但在不同年龄及病理类型间比较,差异无统计学意义(P>0.05)。宫颈癌患者组织中KIF11与β-catenin的表达呈正相关(r=0.461,P<0.05),β-catenin与GSK-3β的表达呈负相关(r=-0.692,P<0.05),KIF11与GSK-3β的表达呈负相关(r=-0.336,P<0.05)。KIF11、β-catenin阳性表达患者的平均生存时间短于阴性表达患者,GSK-3β阳性表达患者的平均生存时间长于阴性表达患者。COX回归分析显示,FIGO分期、淋巴结转移、KIF11、β-catenin为宫颈癌患者预后的独立危险因素,GSK-3β为独立保护因素。结论KIF11、β-catenin、GSK-3β在宫颈癌患者组织中异常表达,KIF11可能参与宫颈癌发生、发展中Wnt/β-catenin通路的调节,三者联合检测可为宫颈癌的诊断及预后提供新的参考。展开更多
基金supported by the National Natural Science Foundation of China(No.82104977)Youth Start-up Foundation of Changhai Hospital(No.2019QNB05)。
文摘Objective:Physical exercise,a common non-drug intervention,is an important strategy in cancer treatment,including hepatocellular carcinoma(HCC).However,the mechanism remains largely unknown.Due to the importance of hypoxia and cancer stemness in the development of HCC,the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness.Methods:A physical exercise intervention of swimming(30 min/d,5 d/week,for 4 weeks)was administered to BALB/c nude mice bearing subcutaneous human HCC tumor.The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring,hematoxylin and eosin(HE)staining,and immunohistochemistry(IHC)detection of proliferating cell nuclear antigen(PCNA)and Ki67.The expression of stemness transcription factors,including Nanog homeobox(NANOG),octamer-binding transcription factor 4(OCT-4),v-Myc avian myelocytomatosis viral oncogene homolog(C-MYC)and hypoxia-inducible factor-1a(HIF-1a),was detected using real-time reverse transcription polymerase chain reaction.A hypoxia probe was used to explore the intratumoral hypoxia status.Western blot was used to detect the expression of HIF-1a and proteins related to protein kinase B(Akt)/glycogen synthase kinase-3β(GSK-3β)/β-catenin signaling pathway.The IHC analysis of platelet endothelial cell adhesion molecule-1(CD31),and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion.SMMC-7721 cells were treated with nude mice serum.The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors.The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1a.Further,the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected.Results:Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor.HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention.Swimming potently alleviated the intratumoral hypoxia,attenuated the cancer stemness,and inhibited the Akt/GSK-3β/β-catenin signaling pathway.Additionally,the desmin+/CD31+ratio,rather than the number of CD31+vessels,was significantly increased in swimming-treated mice.In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere,as well as the m RNA expression level of stemness transcription factors.Consistent with the in vivo results,HIF-1a and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group.Conclusion:Swimming alleviated hypoxia and attenuated cancer stemness in HCC,through suppression of the Akt/GSK-3β/β-catenin signaling pathway.The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor.
文摘目的探讨驱动蛋白家族成员11(kinesin family member 11,KIF11)、β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)在宫颈癌中的表达情况及临床意义。方法采用免疫组化法检测KIF11、β-catenin、GSK-3β在102例宫颈癌、52例高级别鳞状上皮内病变(high-grade squamous intraepithelial lesion,HSIL)、46例低级别鳞状上皮内病变(low-grade squamous intraepithelial lesion,LSIL)及40例慢性宫颈炎组织中的表达情况,分析三者的表达与宫颈癌患者临床病理特征的关系,分析三者之间的相关性,COX比例风险模型分析影响宫颈癌患者预后的影响因素。结果随着宫颈病变进展,KIF11、β-catenin阳性率逐渐升高,GSK-3β阳性率逐渐减低(P<0.05)。KIF11、β-catenin、GSK-3β在宫颈癌组织中的阳性表达在国际妇产科联盟(International Federation of Gynecology and Obstetrics,FIGO)分期、分化程度、淋巴结转移方面比较,差异有统计学意义(P<0.05),但在不同年龄及病理类型间比较,差异无统计学意义(P>0.05)。宫颈癌患者组织中KIF11与β-catenin的表达呈正相关(r=0.461,P<0.05),β-catenin与GSK-3β的表达呈负相关(r=-0.692,P<0.05),KIF11与GSK-3β的表达呈负相关(r=-0.336,P<0.05)。KIF11、β-catenin阳性表达患者的平均生存时间短于阴性表达患者,GSK-3β阳性表达患者的平均生存时间长于阴性表达患者。COX回归分析显示,FIGO分期、淋巴结转移、KIF11、β-catenin为宫颈癌患者预后的独立危险因素,GSK-3β为独立保护因素。结论KIF11、β-catenin、GSK-3β在宫颈癌患者组织中异常表达,KIF11可能参与宫颈癌发生、发展中Wnt/β-catenin通路的调节,三者联合检测可为宫颈癌的诊断及预后提供新的参考。
