Callinice D Capo-Chichi, F. Gnangnon, Charles Kuassi Mekpossi, Sara Hounguè, Xiang-Xi Xu, J. Olory-Togbe
{"title":"组蛋白去乙酰化酶抑制剂在乳腺癌细胞中下调GATA3和ERα需要Lamin A的存在","authors":"Callinice D Capo-Chichi, F. Gnangnon, Charles Kuassi Mekpossi, Sara Hounguè, Xiang-Xi Xu, J. Olory-Togbe","doi":"10.12691/jcrt-6-3-1","DOIUrl":null,"url":null,"abstract":"Background: Breast cancer treatment is challenging due to the inconsistence in tumour biomarker expression including progesterone receptor (PR), estrogen receptor-α (ERα) and GATA3 transcription factor. GATA3 has role in epithelial cell differentiation along with nuclear envelope protein lamin A. The breast cancer cell line MCF7 expresses ERα, abnormal GATA3 isoforms, low PR but lacks lamin A. MCF7 cells are resistant to tamoxifen targeting ERα and more anticancer drugs are being studied to kill them. One of the mechanisms surrounding breast cancer initiation is histone deacetylation. Our objective is to investigate the effect of histone deacetylase inhibitor (HDACI) on PR, ERα and GATA3 expression along with the induction of apoptosis in MCF7 cells forced expressing lamin A. Subsequently, they are also explored as biomarkers for breast cancer prognostic and indicators for targeted breast cancer therapy. Methods: The HDACI used here is Suberoyl-Bis-Hydroxamic Acid (SBHA). Western blot was used to analyze the expression of PR, ERα, and GATA3 in MCF7 control transfected with histone H2B-GFP (MCF7-H2B-GFP) and in MCF7 transfected with lamin A-RFP (MCF7-LA-RFP). The analyses were carried out before and after treatment with DMSO (mock) or SBHA (1 or 2 µM) for 12h. The in vivo expression of the PR, ERα and GATA3 were also explored in 36 archived cell lysates derived from breast cancer micro-biopsies. Results: MCF7-H2B-GFP treated with SBHA increased PR, ERα while MCF7-LA-RFP treated with SBHA reduced ERα and GATA3 but not PR. Biomarkers analysis in ductal carcinoma micro-biopsies derived samples showed that 30% had lost lamin A while 64% expressed PR, ERα and GATA3. Conclusion: In presence of lamin A, SBHA downregulated cancer initiators GATA3 and ERα while inducing cell death. These biomarkers could be useful molecular tools prior to initiating targeted breast cancer therapy.","PeriodicalId":22619,"journal":{"name":"The Journal of Cancer Research","volume":"36 1","pages":"60-69"},"PeriodicalIF":0.0000,"publicationDate":"2018-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Presence of Lamin A is Required for GATA3 and ERα Downregulation by Histone Deacetylase Inhibitor in Breast Cancer Cells\",\"authors\":\"Callinice D Capo-Chichi, F. Gnangnon, Charles Kuassi Mekpossi, Sara Hounguè, Xiang-Xi Xu, J. Olory-Togbe\",\"doi\":\"10.12691/jcrt-6-3-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Background: Breast cancer treatment is challenging due to the inconsistence in tumour biomarker expression including progesterone receptor (PR), estrogen receptor-α (ERα) and GATA3 transcription factor. GATA3 has role in epithelial cell differentiation along with nuclear envelope protein lamin A. The breast cancer cell line MCF7 expresses ERα, abnormal GATA3 isoforms, low PR but lacks lamin A. MCF7 cells are resistant to tamoxifen targeting ERα and more anticancer drugs are being studied to kill them. One of the mechanisms surrounding breast cancer initiation is histone deacetylation. Our objective is to investigate the effect of histone deacetylase inhibitor (HDACI) on PR, ERα and GATA3 expression along with the induction of apoptosis in MCF7 cells forced expressing lamin A. Subsequently, they are also explored as biomarkers for breast cancer prognostic and indicators for targeted breast cancer therapy. Methods: The HDACI used here is Suberoyl-Bis-Hydroxamic Acid (SBHA). Western blot was used to analyze the expression of PR, ERα, and GATA3 in MCF7 control transfected with histone H2B-GFP (MCF7-H2B-GFP) and in MCF7 transfected with lamin A-RFP (MCF7-LA-RFP). The analyses were carried out before and after treatment with DMSO (mock) or SBHA (1 or 2 µM) for 12h. The in vivo expression of the PR, ERα and GATA3 were also explored in 36 archived cell lysates derived from breast cancer micro-biopsies. Results: MCF7-H2B-GFP treated with SBHA increased PR, ERα while MCF7-LA-RFP treated with SBHA reduced ERα and GATA3 but not PR. Biomarkers analysis in ductal carcinoma micro-biopsies derived samples showed that 30% had lost lamin A while 64% expressed PR, ERα and GATA3. Conclusion: In presence of lamin A, SBHA downregulated cancer initiators GATA3 and ERα while inducing cell death. 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引用次数: 0
摘要
背景:由于孕激素受体(PR)、雌激素受体-α (ERα)和GATA3转录因子等肿瘤生物标志物的表达不一致,乳腺癌的治疗具有挑战性。乳腺癌细胞系MCF7表达ERα、异常的GATA3亚型、低PR但缺乏lamin a。MCF7细胞对靶向ERα的他莫昔芬具有耐药性,更多的抗癌药物正在研究中以杀死它们。乳腺癌起始的机制之一是组蛋白去乙酰化。我们的目的是研究组蛋白去乙酰化酶抑制剂(HDACI)在强迫表达lamin a的MCF7细胞中对PR、ERα和GATA3表达的影响,并诱导细胞凋亡。随后,它们也被探索作为乳腺癌预后的生物标志物和乳腺癌靶向治疗的指标。方法:本研究使用的HDACI为亚甲基双羟肟酸(SBHA)。Western blot检测转染组蛋白H2B-GFP的MCF7对照组(MCF7-H2B-GFP)和转染lamin A-RFP的MCF7组(MCF7- la - rfp)中PR、ERα和GATA3的表达情况。在DMSO(模拟)或SBHA(1或2µM)处理12h前后进行分析。我们还在36例乳腺癌显微活检的细胞裂解物中探讨了PR、ERα和GATA3的体内表达。结果:SBHA处理的MCF7-H2B-GFP增加了PR、ERα,而SBHA处理的MCF7-LA-RFP降低了ERα和GATA3,但没有降低PR。导管癌显微活检样本的生物标志物分析显示,30%的细胞失去了层粘胶蛋白A, 64%的细胞表达了PR、ERα和GATA3。结论:在层粘连蛋白A存在的情况下,SBHA可下调肿瘤启动子GATA3和ERα,诱导细胞死亡。这些生物标志物可能是开始靶向乳腺癌治疗之前有用的分子工具。
Presence of Lamin A is Required for GATA3 and ERα Downregulation by Histone Deacetylase Inhibitor in Breast Cancer Cells
Background: Breast cancer treatment is challenging due to the inconsistence in tumour biomarker expression including progesterone receptor (PR), estrogen receptor-α (ERα) and GATA3 transcription factor. GATA3 has role in epithelial cell differentiation along with nuclear envelope protein lamin A. The breast cancer cell line MCF7 expresses ERα, abnormal GATA3 isoforms, low PR but lacks lamin A. MCF7 cells are resistant to tamoxifen targeting ERα and more anticancer drugs are being studied to kill them. One of the mechanisms surrounding breast cancer initiation is histone deacetylation. Our objective is to investigate the effect of histone deacetylase inhibitor (HDACI) on PR, ERα and GATA3 expression along with the induction of apoptosis in MCF7 cells forced expressing lamin A. Subsequently, they are also explored as biomarkers for breast cancer prognostic and indicators for targeted breast cancer therapy. Methods: The HDACI used here is Suberoyl-Bis-Hydroxamic Acid (SBHA). Western blot was used to analyze the expression of PR, ERα, and GATA3 in MCF7 control transfected with histone H2B-GFP (MCF7-H2B-GFP) and in MCF7 transfected with lamin A-RFP (MCF7-LA-RFP). The analyses were carried out before and after treatment with DMSO (mock) or SBHA (1 or 2 µM) for 12h. The in vivo expression of the PR, ERα and GATA3 were also explored in 36 archived cell lysates derived from breast cancer micro-biopsies. Results: MCF7-H2B-GFP treated with SBHA increased PR, ERα while MCF7-LA-RFP treated with SBHA reduced ERα and GATA3 but not PR. Biomarkers analysis in ductal carcinoma micro-biopsies derived samples showed that 30% had lost lamin A while 64% expressed PR, ERα and GATA3. Conclusion: In presence of lamin A, SBHA downregulated cancer initiators GATA3 and ERα while inducing cell death. These biomarkers could be useful molecular tools prior to initiating targeted breast cancer therapy.