Angiotensin-(1-7) [Ang-(1-7)] is a heptapeptide of the renin-angiotensin system with vasodilator and anti-proliferative properties. In the present study, we aim to investigate whether Ang-(1-7) induces apoptosis in br...Angiotensin-(1-7) [Ang-(1-7)] is a heptapeptide of the renin-angiotensin system with vasodilator and anti-proliferative properties. In the present study, we aim to investigate whether Ang-(1-7) induces apoptosis in breast cancer cells and whether the altered expression of apoptosis-related genes is involved in this process. Human breast cell line T47D was treated with angiotensin-(1-7) and angiotensin II (Ang II). Cell proliferation and apoptosis were quantified using hemocytometer and flow cytometry, respectively. The expression of 84 apoptosis-related genes was evaluated through qPCR array. Ang-(1-7), as opposed to Ang II, decreased proliferation and increased apoptosis in T47D cells. Moreover, many pro-apoptotic genes were up-regulated, such as BAK1, BAX, BCL2L1, BID and BIK. In addition, some anti-apoptotic genes as AKT1 and XIAP were down-regulated by heptapeptide. Although a deeper study should be performed, our results support the hypothesis that Ang-(1-7) could change the expression of several genes related to apoptosis, interfering directly in the molecular pathways associated with the survival of breast cancer cells.展开更多
Breast cancer is the most common cancer among women. Angiotensin-(1 - 7) [Ang-(1 - 7)] has been correlated with cancer antiproliferative and apoptotic effects, similar properties of the human Chorionic Gonadotrofin (h...Breast cancer is the most common cancer among women. Angiotensin-(1 - 7) [Ang-(1 - 7)] has been correlated with cancer antiproliferative and apoptotic effects, similar properties of the human Chorionic Gonadotrofin (hCG). The aims of this work are to evaluate the role of Ang-(1 - 7) and of hCG in modulating the expression of Nuclear Receptors and Coregulators related genes in the tumorigenic breast cell line SK-BR3. Three experimental groups were created: control, hCG and hCG + Ang-(1 - 7). Cells were treated for 11 days and then had their RNA extracted. Samples were loaded into PCR Array plates containing 84 genes relate to Nuclear Receptors and Coregulators pathways. Gene expression data were used to construct canonical pathways (MetacoreTM). hCG and hCG + Ang-(1 - 7) treatments markedly modulate the expression of Nuclear Receptors and Coregulators related genes. hCG differentially expressed 17% of the genes, being 29% upregulated and 71% downregulated. Meanwhile, hCG + Ang-(1 - 7) changed the expression of 30% of the genes on the plate, among these genes 56% were upregulated and 44% downregulated. Among these differentially expressed genes, we highlight Esr1, Nr2f2, and Nr2f1, Esr1, Hdac5, and Nr4A1 (>4 fold). Finally MetaCore analysis based on Gene Ontology (GO) generated six networks for hCG and ten networks for the combined treatment. All generated networks are related to regulation of apoptosis or to Programmed Cell Death processes. In summary, our results herein demonstrate that the modulation of sexual hormones and of other nuclear factor genes expression might underlie the tumorigenic protection effect and the induction of cell differentiation caused by the hormones hCG and Ang-(1 - 7), especially in Cancer Stem Cells.展开更多
目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling...目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling）法检测CIK细胞的凋亡.通过免疫细胞化学染色法检测ZK-75-1细胞中p53、p16、C-myc、Bcl-2及Bax的表达率.结果：HE染色显示,CIK细胞向ZK-75-1细胞靠近,形成典型的玫瑰花环状;肿瘤细胞的胞浆中出现颗粒状物,有的肿瘤细胞只见颗粒状碎片;而作为对照的乳腺癌细胞生长良好.TUNEL法检测显示,对照组细胞未染色或染呈均匀的淡蓝色;实验组凋亡的细胞缩小,核或核周染呈深蓝色.CIK细胞作用4～12 h ZK-75-1细胞的凋亡率上升,作用12～24h细胞的凋亡率下降,与对照组相比较有统计学意义（P＜0.01）.免疫细胞化学染色的结果表明,CIK细胞实验组p53、pi6、C-myc及Bcl-2蛋白随作用时间的延长均下降,Bax蛋白的表达上调,与对照组相比较均有统计学意义（P＜0.01）.结论：CIK细胞对乳腺癌细胞ZK-75-1杀伤作用的机制之一,可能与p53、p16、C-myc、Bcl-2蛋白表达的下调及Bax蛋白表达的上调有关,并与CIK细胞作用的时间关系密切.展开更多
MiR-142-3p has been reported to act as a tumor suppressor in breast cancer.However,the regulatory effect of miR-142-3p on drug resistance of breast cancer cells and its underlying mechanism remain unknown.Here,we foun...MiR-142-3p has been reported to act as a tumor suppressor in breast cancer.However,the regulatory effect of miR-142-3p on drug resistance of breast cancer cells and its underlying mechanism remain unknown.Here,we found that miR-142-3p was significantly downregulated in the doxorubicin(DOX)-resistant MCF-7 cell line(MCF-7/DOX).MiR-142-3p overexpression increased DOX sensitivity and enhanced DOXinduced apoptosis in breast cancer cells.High-mobility group box 1(HMGB1)is a direct functional target of miR-142-3p in breast cancer cells and miR-142-3p negatively regulated HMGB1 expression.Moreover,overexpres sion of HMGB1 dramatically reversed the promotion of apoptosis and inhibition of autophagy mediated by miR-142-3p up-regulation.In conclusion,miR-142-3p overexpression may inhibit autophagy and promote the drug sensitivity of breast cancer cells to DOX by targeting HMGB 1.The miR-142-3 p/HMGB1 axis might be a novel target to regulate the drug resistance of breast cancer patients.展开更多
基金supported by grants number 2008/54383-0,2010/03658-9 and 2011/08531-0 from the Sao Paulo Research Foundation(FAPESP)-Brazil.
文摘Angiotensin-(1-7) [Ang-(1-7)] is a heptapeptide of the renin-angiotensin system with vasodilator and anti-proliferative properties. In the present study, we aim to investigate whether Ang-(1-7) induces apoptosis in breast cancer cells and whether the altered expression of apoptosis-related genes is involved in this process. Human breast cell line T47D was treated with angiotensin-(1-7) and angiotensin II (Ang II). Cell proliferation and apoptosis were quantified using hemocytometer and flow cytometry, respectively. The expression of 84 apoptosis-related genes was evaluated through qPCR array. Ang-(1-7), as opposed to Ang II, decreased proliferation and increased apoptosis in T47D cells. Moreover, many pro-apoptotic genes were up-regulated, such as BAK1, BAX, BCL2L1, BID and BIK. In addition, some anti-apoptotic genes as AKT1 and XIAP were down-regulated by heptapeptide. Although a deeper study should be performed, our results support the hypothesis that Ang-(1-7) could change the expression of several genes related to apoptosis, interfering directly in the molecular pathways associated with the survival of breast cancer cells.
基金supported by Grant number 2011/10516-0 and 2008/54383-0 from the Sao Paulo Research Foundation(FAPESP)-Brazil.
文摘Breast cancer is the most common cancer among women. Angiotensin-(1 - 7) [Ang-(1 - 7)] has been correlated with cancer antiproliferative and apoptotic effects, similar properties of the human Chorionic Gonadotrofin (hCG). The aims of this work are to evaluate the role of Ang-(1 - 7) and of hCG in modulating the expression of Nuclear Receptors and Coregulators related genes in the tumorigenic breast cell line SK-BR3. Three experimental groups were created: control, hCG and hCG + Ang-(1 - 7). Cells were treated for 11 days and then had their RNA extracted. Samples were loaded into PCR Array plates containing 84 genes relate to Nuclear Receptors and Coregulators pathways. Gene expression data were used to construct canonical pathways (MetacoreTM). hCG and hCG + Ang-(1 - 7) treatments markedly modulate the expression of Nuclear Receptors and Coregulators related genes. hCG differentially expressed 17% of the genes, being 29% upregulated and 71% downregulated. Meanwhile, hCG + Ang-(1 - 7) changed the expression of 30% of the genes on the plate, among these genes 56% were upregulated and 44% downregulated. Among these differentially expressed genes, we highlight Esr1, Nr2f2, and Nr2f1, Esr1, Hdac5, and Nr4A1 (>4 fold). Finally MetaCore analysis based on Gene Ontology (GO) generated six networks for hCG and ten networks for the combined treatment. All generated networks are related to regulation of apoptosis or to Programmed Cell Death processes. In summary, our results herein demonstrate that the modulation of sexual hormones and of other nuclear factor genes expression might underlie the tumorigenic protection effect and the induction of cell differentiation caused by the hormones hCG and Ang-(1 - 7), especially in Cancer Stem Cells.
文摘目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling）法检测CIK细胞的凋亡.通过免疫细胞化学染色法检测ZK-75-1细胞中p53、p16、C-myc、Bcl-2及Bax的表达率.结果：HE染色显示,CIK细胞向ZK-75-1细胞靠近,形成典型的玫瑰花环状;肿瘤细胞的胞浆中出现颗粒状物,有的肿瘤细胞只见颗粒状碎片;而作为对照的乳腺癌细胞生长良好.TUNEL法检测显示,对照组细胞未染色或染呈均匀的淡蓝色;实验组凋亡的细胞缩小,核或核周染呈深蓝色.CIK细胞作用4～12 h ZK-75-1细胞的凋亡率上升,作用12～24h细胞的凋亡率下降,与对照组相比较有统计学意义（P＜0.01）.免疫细胞化学染色的结果表明,CIK细胞实验组p53、pi6、C-myc及Bcl-2蛋白随作用时间的延长均下降,Bax蛋白的表达上调,与对照组相比较均有统计学意义（P＜0.01）.结论：CIK细胞对乳腺癌细胞ZK-75-1杀伤作用的机制之一,可能与p53、p16、C-myc、Bcl-2蛋白表达的下调及Bax蛋白表达的上调有关,并与CIK细胞作用的时间关系密切.
基金the financial support by National Natural Science Foundation of China(Nos.81330007 and U1601227)the Science and Technology Programs of Guangdong Province(Nos.2014A050503047 and 2015B020225006,China)National Natural Science Foundation of China(81700382)
文摘MiR-142-3p has been reported to act as a tumor suppressor in breast cancer.However,the regulatory effect of miR-142-3p on drug resistance of breast cancer cells and its underlying mechanism remain unknown.Here,we found that miR-142-3p was significantly downregulated in the doxorubicin(DOX)-resistant MCF-7 cell line(MCF-7/DOX).MiR-142-3p overexpression increased DOX sensitivity and enhanced DOXinduced apoptosis in breast cancer cells.High-mobility group box 1(HMGB1)is a direct functional target of miR-142-3p in breast cancer cells and miR-142-3p negatively regulated HMGB1 expression.Moreover,overexpres sion of HMGB1 dramatically reversed the promotion of apoptosis and inhibition of autophagy mediated by miR-142-3p up-regulation.In conclusion,miR-142-3p overexpression may inhibit autophagy and promote the drug sensitivity of breast cancer cells to DOX by targeting HMGB 1.The miR-142-3 p/HMGB1 axis might be a novel target to regulate the drug resistance of breast cancer patients.
