目的明确泛素特异性蛋白酶41(ubiguitin-specific peptidase 41,USP41)在三阴性乳腺癌(triple-negative breast cancer,TNBC)的表达情况,及其与TNBC恶性表型及阿霉素敏感性的相关性和潜在作用机制。方法采用Western blot、qPCR检测USP41...目的明确泛素特异性蛋白酶41(ubiguitin-specific peptidase 41,USP41)在三阴性乳腺癌(triple-negative breast cancer,TNBC)的表达情况,及其与TNBC恶性表型及阿霉素敏感性的相关性和潜在作用机制。方法采用Western blot、qPCR检测USP41在TNBC耐药细胞株及临床组织样本的表达情况。随后确定USP41分子高表达,并通过CCK8、克隆形成实验、Transwell、Western blot及CoIP-MS等细胞生物学方法评价USP41在TNBC恶性表型及阿霉素耐药中的作用及机制。结果USP41在TNBC样本的表达显著高于癌旁组织。USP41在阿霉素耐药细胞株MDA-MB-231/DXR中表达量高出近40倍,IC50值为6.86μM。干扰USP41可显著增加MDA-MB-231/DXR细胞对阿霉素的敏感性。干扰USP41可显著的抑制细胞的细胞增殖、克隆形成及迁移能力,克隆数量降低30%~80%,迁移细胞数量降低超过70%,差异有统计学意义。此外,USP41敲低可提高MDA-MB-231细胞对阿霉素的敏感性,IC50由5.49μM降低到2.36μM和2.56μM。CO-IP结果显示USP41可与活化的蛋白激酶C1受体(receptor of activated protein kinase C1,RACK1)直接相互作用,且RACK1在癌组织的表达显著高于癌旁。干扰RACK1可抑制MDA-MB-231细胞增殖,IC50由9.87μM降低到4.67μM和4.36μM。克隆形成能力下降超过30%,差异有统计学意义。相较于对照组,USP41敲减使细胞迁移能力下降超过70%。结论USP41高表达与TNBC恶性表型及阿霉素耐药相关,且RACK1可能是USP41发挥作用的关键分子。展开更多
Breast cancer is the most frequently diagnosed cancer in women,accounting for 30%of new diagnosing female cancers.Emerging evidence suggests that ubiquitin and ubiquitination played a role in a number of breast cancer...Breast cancer is the most frequently diagnosed cancer in women,accounting for 30%of new diagnosing female cancers.Emerging evidence suggests that ubiquitin and ubiquitination played a role in a number of breast cancer etiology and progression processes.As the primary deubiquitinases in the family,ubiquitin-specific peptidases(USPs)are thought to represent potential therapeutic targets.The role of ubiquitin and ubiquitination in breast cancer,as well as the classification and involvement of USPs are discussed in this review,such as USP1,USP4,USP7,USP9X,USP14,USP18,USP20,USP22,USP25,USP37,and USP39.The reported USPs inhibitors investigated in breast cancer were also summarized,along with the signaling pathways involved in the investigation and its study phase.Despite no USP inhibitor has yet been approved for clinical use,the biological efficacy indicated their potential in breast cancer treatment.With the improvements in phenotypic discovery,we will know more about USPs and USPs inhibitors,developing more potent and selective clinical candidates for breast cancer.展开更多
目的探索USP41在乳腺癌组织样本及MCF7乳腺癌细胞中的表达情况,及其与恶性表型的相关性和潜在作用机制。方法采用Western blot法、qPCR法检测USP41在MCF7细胞株及临床组织样本中的表达情况。随后通过CCK-8、克隆形成实验、Transwell、We...目的探索USP41在乳腺癌组织样本及MCF7乳腺癌细胞中的表达情况,及其与恶性表型的相关性和潜在作用机制。方法采用Western blot法、qPCR法检测USP41在MCF7细胞株及临床组织样本中的表达情况。随后通过CCK-8、克隆形成实验、Transwell、Western blot以及CoIP-MS等细胞生物学方法评价USP41对MCF7的作用及机制。结果USP41在乳腺癌样本及MCF7细胞株的表达显著高于癌旁组织。干扰USP41可以显著地抑制细胞的增殖、克隆形成及迁移能力,促进细胞凋亡。CoIP结果显示,USP41可以与活化蛋白激酶C受体1(receptor for activated C kinase 1,RACK1)直接相互作用,且RACK1在癌组织的表达显著高于癌旁。干扰RACK1可抑制MCF7细胞增殖、克隆形成及迁移能力。结论USP41在MCF7中高表达,且通过上调RACK1促进MCF7乳腺癌细胞增殖、迁移,抑制细胞凋亡。展开更多
文摘目的明确泛素特异性蛋白酶41(ubiguitin-specific peptidase 41,USP41)在三阴性乳腺癌(triple-negative breast cancer,TNBC)的表达情况,及其与TNBC恶性表型及阿霉素敏感性的相关性和潜在作用机制。方法采用Western blot、qPCR检测USP41在TNBC耐药细胞株及临床组织样本的表达情况。随后确定USP41分子高表达,并通过CCK8、克隆形成实验、Transwell、Western blot及CoIP-MS等细胞生物学方法评价USP41在TNBC恶性表型及阿霉素耐药中的作用及机制。结果USP41在TNBC样本的表达显著高于癌旁组织。USP41在阿霉素耐药细胞株MDA-MB-231/DXR中表达量高出近40倍,IC50值为6.86μM。干扰USP41可显著增加MDA-MB-231/DXR细胞对阿霉素的敏感性。干扰USP41可显著的抑制细胞的细胞增殖、克隆形成及迁移能力,克隆数量降低30%~80%,迁移细胞数量降低超过70%,差异有统计学意义。此外,USP41敲低可提高MDA-MB-231细胞对阿霉素的敏感性,IC50由5.49μM降低到2.36μM和2.56μM。CO-IP结果显示USP41可与活化的蛋白激酶C1受体(receptor of activated protein kinase C1,RACK1)直接相互作用,且RACK1在癌组织的表达显著高于癌旁。干扰RACK1可抑制MDA-MB-231细胞增殖,IC50由9.87μM降低到4.67μM和4.36μM。克隆形成能力下降超过30%,差异有统计学意义。相较于对照组,USP41敲减使细胞迁移能力下降超过70%。结论USP41高表达与TNBC恶性表型及阿霉素耐药相关,且RACK1可能是USP41发挥作用的关键分子。
基金Supported by the National Natural Science Foundation of China,No.81472598Project of Xijing Hospital,No.XJZT18MJ30.
文摘Breast cancer is the most frequently diagnosed cancer in women,accounting for 30%of new diagnosing female cancers.Emerging evidence suggests that ubiquitin and ubiquitination played a role in a number of breast cancer etiology and progression processes.As the primary deubiquitinases in the family,ubiquitin-specific peptidases(USPs)are thought to represent potential therapeutic targets.The role of ubiquitin and ubiquitination in breast cancer,as well as the classification and involvement of USPs are discussed in this review,such as USP1,USP4,USP7,USP9X,USP14,USP18,USP20,USP22,USP25,USP37,and USP39.The reported USPs inhibitors investigated in breast cancer were also summarized,along with the signaling pathways involved in the investigation and its study phase.Despite no USP inhibitor has yet been approved for clinical use,the biological efficacy indicated their potential in breast cancer treatment.With the improvements in phenotypic discovery,we will know more about USPs and USPs inhibitors,developing more potent and selective clinical candidates for breast cancer.
文摘目的探索USP41在乳腺癌组织样本及MCF7乳腺癌细胞中的表达情况,及其与恶性表型的相关性和潜在作用机制。方法采用Western blot法、qPCR法检测USP41在MCF7细胞株及临床组织样本中的表达情况。随后通过CCK-8、克隆形成实验、Transwell、Western blot以及CoIP-MS等细胞生物学方法评价USP41对MCF7的作用及机制。结果USP41在乳腺癌样本及MCF7细胞株的表达显著高于癌旁组织。干扰USP41可以显著地抑制细胞的增殖、克隆形成及迁移能力,促进细胞凋亡。CoIP结果显示,USP41可以与活化蛋白激酶C受体1(receptor for activated C kinase 1,RACK1)直接相互作用,且RACK1在癌组织的表达显著高于癌旁。干扰RACK1可抑制MCF7细胞增殖、克隆形成及迁移能力。结论USP41在MCF7中高表达,且通过上调RACK1促进MCF7乳腺癌细胞增殖、迁移,抑制细胞凋亡。
