柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)最早作为2型和5型腺病毒的受体而被人们发现和认识。大量研究发现CAR可以影响肿瘤细胞的生长、细胞骨架的变化、细胞间的粘附等,从而在肿瘤侵袭转移过程中起非常重要的作用。...柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)最早作为2型和5型腺病毒的受体而被人们发现和认识。大量研究发现CAR可以影响肿瘤细胞的生长、细胞骨架的变化、细胞间的粘附等,从而在肿瘤侵袭转移过程中起非常重要的作用。而且,CAR的表达与肿瘤的预后和腺病毒介导的减瘤效应也有密切的关系。基于CAR的重要作用,CAR已逐渐成为肿瘤发生发展机制及治疗领域研究中新的"热点",本文结合CAR的基本结构及功能对以上研究的进展作一综述。展开更多
目的以柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)蛋白结构的功能解析作为切入点,构建不同CAR胞内域缺失突变的表达载体。方法利用定点缺失突变策略构建CAR胞内域PKC、CK2、TYR磷酸化位点的缺失突变体及胞内域完全缺失...目的以柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)蛋白结构的功能解析作为切入点,构建不同CAR胞内域缺失突变的表达载体。方法利用定点缺失突变策略构建CAR胞内域PKC、CK2、TYR磷酸化位点的缺失突变体及胞内域完全缺失的突变体。利用三步PCR法扩增获得相关的基因片段后,将其克隆入真核表达载体pcDNA3.1/V5/His,通过双酶切及测序进行鉴定后,分别转染至低表达CAR的人卵巢癌细胞系SKOV3细胞中,并用Werstern blot检测细胞转染后CAR的表达水平。结果所有突变体经过测序分析均证实序列与设想的突变序列相符,各突变体在SKOV3细胞中获得了满意的表达。结论成功构建不同CAR胞内功能域缺失突变表达载体,为后续研究CAR及其胞内不同功能域在卵巢癌中的作用机制提供了研究基础。展开更多
目的观察转染转移相关基因1(metastasis-associated 1,MTA1)基因对人宫颈癌细胞HeLa迁移、侵袭、粘附能力以及β-catenin、基质金属蛋白酶-9(MMP-9)表达的影响。方法 Western blot法检测HeLa、SiHa细胞中MTA1表达;以脂质体Lipofectamine...目的观察转染转移相关基因1(metastasis-associated 1,MTA1)基因对人宫颈癌细胞HeLa迁移、侵袭、粘附能力以及β-catenin、基质金属蛋白酶-9(MMP-9)表达的影响。方法 Western blot法检测HeLa、SiHa细胞中MTA1表达;以脂质体LipofectamineTM2000介导的方法将含MTA1全长基因的质粒pEGFP-C1-MTA1转染HeLa细胞,Western blot法检测目的基因表达;划痕、侵袭、粘附试验分别检测HeLa细胞迁移、侵袭、粘附能力变化;Western blot法检测β-catenin、MMP-9表达变化。结果与SiHa细胞相比,MTA1在HeLa细胞中表达较低(P<0.05);MTA1质粒转染的HeLa细胞中,MTA1表达、划痕愈合、Matrigel侵袭、Matrigel粘附能力均明显高于未转染及转染空载体对照组(均P<0.05);β-catenin、MMP-9表达在转染后均上调(均P<0.05)。结论 MTA1基因过表达可促进HeLa细胞转移、侵袭和粘附,上调β-catenin、MMP-9表达,MTA1基因可能成为肿瘤治疗靶点。展开更多
It has been reported that metastasis-associated gene 1 （Mta1） is overexpressed in many malignant tumors with high metastatic potential. In addition, some studies indicated that MTA1 participated in invasion, metasta...It has been reported that metastasis-associated gene 1 （Mta1） is overexpressed in many malignant tumors with high metastatic potential. In addition, some studies indicated that MTA1 participated in invasion, metastasis, and survival of cancer cells by regulating cell migration, adhesion and proliferation. But the role of MTA1 is unclear in vitro in the development of cervical cancer cells. This study investigated whether and how MTA1 mediated cell proliferation, migration, invasion and adhesion in cervical cancer. MTA1 expression level was detected by Western blot in two cervical cancer cell lines of different invasion potentials. The effects of MTA1 expression on SiHa cell apoptosis, cycle, proliferation, migration, invasion and adhesion were tested by flow cytometry, MTT, wound-healing assay, Transwell assay and adhesion assay, respectively. The expression levels of p53, E-cadherin, and β-catenin activity were evaluated in untreated and treated cells. The results showed that MTA1 protein expression was significantly higher in SiHa than in HeLa, which was correlated well with the potential of migration and invasion in both cell lines. Furthermore, the cell invasion, migration and adhesion capabilities were decreased after inhibition of MTA1 expression mediated by Mta1-siRNA transfection in SiHa. However, no significant differences were found in cell apoptosis, cycle, and proliferation. In addition, E-cadherin and p53 protein levels were significantly up-regulated, while β-catenin was significantly down-regulated in SiHa transfected with the siRNA. These results demonstrated that MTA1 played an important role in the migration and invasion of cervical cancer cells. It was speculated that the decreased migration and invasion capability by inhibiting the MTA1 expression in the SiHa cell line may be mediated through the altered expression of p53, and E-cadherin/β-catenin complex. MTA1 could serve as a potential therapeutic target in cervical cancer.展开更多
文摘柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)最早作为2型和5型腺病毒的受体而被人们发现和认识。大量研究发现CAR可以影响肿瘤细胞的生长、细胞骨架的变化、细胞间的粘附等,从而在肿瘤侵袭转移过程中起非常重要的作用。而且,CAR的表达与肿瘤的预后和腺病毒介导的减瘤效应也有密切的关系。基于CAR的重要作用,CAR已逐渐成为肿瘤发生发展机制及治疗领域研究中新的"热点",本文结合CAR的基本结构及功能对以上研究的进展作一综述。
文摘目的以柯萨奇-腺病毒受体(Coxsackie and adenovirus receptor,CAR)蛋白结构的功能解析作为切入点,构建不同CAR胞内域缺失突变的表达载体。方法利用定点缺失突变策略构建CAR胞内域PKC、CK2、TYR磷酸化位点的缺失突变体及胞内域完全缺失的突变体。利用三步PCR法扩增获得相关的基因片段后,将其克隆入真核表达载体pcDNA3.1/V5/His,通过双酶切及测序进行鉴定后,分别转染至低表达CAR的人卵巢癌细胞系SKOV3细胞中,并用Werstern blot检测细胞转染后CAR的表达水平。结果所有突变体经过测序分析均证实序列与设想的突变序列相符,各突变体在SKOV3细胞中获得了满意的表达。结论成功构建不同CAR胞内功能域缺失突变表达载体,为后续研究CAR及其胞内不同功能域在卵巢癌中的作用机制提供了研究基础。
基金supported by grants from the Major State Basic Research Development Program of China (973 Program,No. 2009CB521808)the National Natural Sciences Foundation of China (No. 30700895)
文摘It has been reported that metastasis-associated gene 1 （Mta1） is overexpressed in many malignant tumors with high metastatic potential. In addition, some studies indicated that MTA1 participated in invasion, metastasis, and survival of cancer cells by regulating cell migration, adhesion and proliferation. But the role of MTA1 is unclear in vitro in the development of cervical cancer cells. This study investigated whether and how MTA1 mediated cell proliferation, migration, invasion and adhesion in cervical cancer. MTA1 expression level was detected by Western blot in two cervical cancer cell lines of different invasion potentials. The effects of MTA1 expression on SiHa cell apoptosis, cycle, proliferation, migration, invasion and adhesion were tested by flow cytometry, MTT, wound-healing assay, Transwell assay and adhesion assay, respectively. The expression levels of p53, E-cadherin, and β-catenin activity were evaluated in untreated and treated cells. The results showed that MTA1 protein expression was significantly higher in SiHa than in HeLa, which was correlated well with the potential of migration and invasion in both cell lines. Furthermore, the cell invasion, migration and adhesion capabilities were decreased after inhibition of MTA1 expression mediated by Mta1-siRNA transfection in SiHa. However, no significant differences were found in cell apoptosis, cycle, and proliferation. In addition, E-cadherin and p53 protein levels were significantly up-regulated, while β-catenin was significantly down-regulated in SiHa transfected with the siRNA. These results demonstrated that MTA1 played an important role in the migration and invasion of cervical cancer cells. It was speculated that the decreased migration and invasion capability by inhibiting the MTA1 expression in the SiHa cell line may be mediated through the altered expression of p53, and E-cadherin/β-catenin complex. MTA1 could serve as a potential therapeutic target in cervical cancer.
