The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. An-tisense EphB4 vectors and shRN...The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. An-tisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin （mTOR） were constructed and transfected into A2780 and SKOV3 cells （two ovarian cancer cell lines）. The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.展开更多
TMTP1, a 5-amino acid peptide NVVRQ, obtained by using the flagella peptide library screening in our previous studies, can be used for the labeling of malignant in situ and metastatic lesions, and even micro-metastase...TMTP1, a 5-amino acid peptide NVVRQ, obtained by using the flagella peptide library screening in our previous studies, can be used for the labeling of malignant in situ and metastatic lesions, and even micro-metastases. In this study, TMTP1 was assessed for its ability to specifically target the malignant hematopoietic cells and metastatic lesions of hematological malignancies. FITC-TMTP1 was chemically synthesized. Immunofluorescence assay and competitive test were carried out to determine the specific binding capacity of TMTPl to hematological malignant cell lines, including HL60, k562, SHI-1, Jurkat, Raji, El-4 and umbilical cord blood mononuclear cells. Mononuclear cells were isolated from the bone marrow of healthy subjects and patients with chronic myeloid leukemia. Then the cells were co-clutured with TMTP1 or scrambled peptides and the binding and affinity of TMTP1 peptide to the primary cells of hematological malignancies were flow cytometrically analyzed. The binding speci-ficity of TMTP1 to target hematological malignancies was measured in vivo by intravenous injection of FITC-conjugated TMTP1 into El-4 lymphoma-bearing mice. The results showed that TMTP1 specifi-cally bound to the cells of a series of hematological malignancies, including HL60, k562, Jurkat, Raji , El-4 and chronic myeloid leukemia primary cells but not to bone marrow mononuclear cells from healthy subjects. By contrast, TMTP1 could bind to the metastatic foci of lymphoma originating from the EL-4 cell line while the scrambled peptide failed to do so. Moreover, the occult metastases could be identified, with high specificity, by detecting FITC-TMTP1. We are led to conclude that TMTP1, as a novel tumor-homing peptide, can serve as a marker for primary malignant and metastatic lesions for the early diagnosis of hematological malignances and a carrier of anticancer drugs for cancer treatment.展开更多
目的:研究哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路蛋白在人卵巢上皮性癌中的表达及其临床意义。方法:选取武汉同济医院妇产科2005～2008间手术切除并经病理证实的人卵巢上皮性癌63例,其中Ⅰ级12例、Ⅱ级1...目的:研究哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路蛋白在人卵巢上皮性癌中的表达及其临床意义。方法:选取武汉同济医院妇产科2005～2008间手术切除并经病理证实的人卵巢上皮性癌63例,其中Ⅰ级12例、Ⅱ级15例、Ⅲ级23例、Ⅳ级13例,应用免疫组织化学法检测卵巢上皮性癌组织中mTOR信号通路关键蛋白pm-TOR、pAKT的表达,分析其与卵巢癌患者临床病理特征的关系。结果:pmTOR在各级卵巢上皮性癌中的阳性率分别为:Ⅰ级41.7%(5/12)、Ⅱ级53.3%(8/15)、Ⅲ级65.2%(15/23)、Ⅳ级69.2%(9/13),pAKT在各级卵巢上皮性癌中的阳性率分别为:Ⅰ级33.3%(5/12)、Ⅱ级40.0%(6/15)、Ⅲ级52.2%(12/23)、Ⅳ级53.8%(7/13);各阳性表达率间无统计学差异(P>0.05),但其表达水平均随着卵巢上皮性癌病理级别的增高而升高,而与病理类型无关。结论:mTOR信号通路蛋白在卵巢上皮性癌的发生发展中起重要的作用,可能是卵巢癌治疗的潜在靶点。展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81001006)the"973"Program of China(No.2009CB521800)the Specialized Research Fund for the Doctoral Program of Higher Education of China(No.200804871030)
文摘The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. An-tisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin （mTOR） were constructed and transfected into A2780 and SKOV3 cells （two ovarian cancer cell lines）. The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.
基金supported by the National Science Foundation of China (No. 30800402)
文摘TMTP1, a 5-amino acid peptide NVVRQ, obtained by using the flagella peptide library screening in our previous studies, can be used for the labeling of malignant in situ and metastatic lesions, and even micro-metastases. In this study, TMTP1 was assessed for its ability to specifically target the malignant hematopoietic cells and metastatic lesions of hematological malignancies. FITC-TMTP1 was chemically synthesized. Immunofluorescence assay and competitive test were carried out to determine the specific binding capacity of TMTPl to hematological malignant cell lines, including HL60, k562, SHI-1, Jurkat, Raji, El-4 and umbilical cord blood mononuclear cells. Mononuclear cells were isolated from the bone marrow of healthy subjects and patients with chronic myeloid leukemia. Then the cells were co-clutured with TMTP1 or scrambled peptides and the binding and affinity of TMTP1 peptide to the primary cells of hematological malignancies were flow cytometrically analyzed. The binding speci-ficity of TMTP1 to target hematological malignancies was measured in vivo by intravenous injection of FITC-conjugated TMTP1 into El-4 lymphoma-bearing mice. The results showed that TMTP1 specifi-cally bound to the cells of a series of hematological malignancies, including HL60, k562, Jurkat, Raji , El-4 and chronic myeloid leukemia primary cells but not to bone marrow mononuclear cells from healthy subjects. By contrast, TMTP1 could bind to the metastatic foci of lymphoma originating from the EL-4 cell line while the scrambled peptide failed to do so. Moreover, the occult metastases could be identified, with high specificity, by detecting FITC-TMTP1. We are led to conclude that TMTP1, as a novel tumor-homing peptide, can serve as a marker for primary malignant and metastatic lesions for the early diagnosis of hematological malignances and a carrier of anticancer drugs for cancer treatment.
文摘目的:研究哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路蛋白在人卵巢上皮性癌中的表达及其临床意义。方法:选取武汉同济医院妇产科2005～2008间手术切除并经病理证实的人卵巢上皮性癌63例,其中Ⅰ级12例、Ⅱ级15例、Ⅲ级23例、Ⅳ级13例,应用免疫组织化学法检测卵巢上皮性癌组织中mTOR信号通路关键蛋白pm-TOR、pAKT的表达,分析其与卵巢癌患者临床病理特征的关系。结果:pmTOR在各级卵巢上皮性癌中的阳性率分别为:Ⅰ级41.7%(5/12)、Ⅱ级53.3%(8/15)、Ⅲ级65.2%(15/23)、Ⅳ级69.2%(9/13),pAKT在各级卵巢上皮性癌中的阳性率分别为:Ⅰ级33.3%(5/12)、Ⅱ级40.0%(6/15)、Ⅲ级52.2%(12/23)、Ⅳ级53.8%(7/13);各阳性表达率间无统计学差异(P>0.05),但其表达水平均随着卵巢上皮性癌病理级别的增高而升高,而与病理类型无关。结论:mTOR信号通路蛋白在卵巢上皮性癌的发生发展中起重要的作用,可能是卵巢癌治疗的潜在靶点。
