Cheng Chen, Shan Guo, Wenying Chai, Jun Yang, Ying Yang, Guimin Chen, Haishan Rao, Yun Ma, Song Bai
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ESTIMATE algorithms were utilized to assess immune infiltrative patterns. Prognostic gene expression patterns were derived from LASSO regression and univariate COX regression analysis. Subsequently, these signatures underwent examination via use of the Kaplan-Meier survival curve. 6 pairs of fresh tissue specimens (tumor and adjacent non-tumor) were employed to assess the expression of 7 ARGs genes via qPCR. Notably, DCN and FOS were not expressed in BC tissue, which had been excluded in our subsequent experiments. Also, among remaining 5 ARGs, solely the expression of ADH1A demonstrated a statistically remarkable disparity between freshly collected cancer tissues and the adjacent ones. ADH1A-overexpressed and ADH1A-sh vectors were transfected into MCF-7 and MCF-7-AR cell lines, respectively. The expression status of FABP4, CALML5, ADH1A, C1orf106, CIDEC, β-catenin, N-cadherin, and Vimentin in the clinical samples were scrutinized using RT-qPCR and western blotting techniques. Migration and invasion through transwell chambers were employed to assess the migratory and invasive potential of the cells. Detailed evaluation of cell proliferation was conducted utilizing a Cell Counting Kit-8 (CCK-8) assay. The apoptotic index of the cells was determined by flow cytometry analysis. An innovative anoikis-associated signature consisting of seven genes, namely ADH1A, DCN, CIEDC, FABP4, FOS, CALML5, and C1orf106, was devised to stratify BC patients into high- and low-risk cohorts. This unique risk assessment model, formulated via the distinctive signature approach, has been validated as an independent prognostic indicator. Additional analysis demonstrated that distinct risk subtypes manifested variances in the tumor microenvironment and drug sensitivities. Suppression of ADH1A enhanced the migratory and invasive capacities and reduced these tumorigenesis-related protein levels, underscoring the prognostic role of ADH1A in the progression of BC. 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Subsequently, these signatures underwent examination via use of the Kaplan-Meier survival curve. 6 pairs of fresh tissue specimens (tumor and adjacent non-tumor) were employed to assess the expression of 7 ARGs genes via qPCR. Notably, DCN and FOS were not expressed in BC tissue, which had been excluded in our subsequent experiments. Also, among remaining 5 ARGs, solely the expression of ADH1A demonstrated a statistically remarkable disparity between freshly collected cancer tissues and the adjacent ones. ADH1A-overexpressed and ADH1A-sh vectors were transfected into MCF-7 and MCF-7-AR cell lines, respectively. The expression status of FABP4, CALML5, ADH1A, C1orf106, CIDEC, β-catenin, N-cadherin, and Vimentin in the clinical samples were scrutinized using RT-qPCR and western blotting techniques. Migration and invasion through transwell chambers were employed to assess the migratory and invasive potential of the cells. 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引用次数: 0
摘要
乳腺癌(BC)是一种广泛存在的致命肿瘤,与 BC 的亚型无关。转移仍然是导致不良预后的关键因素。确定新的诊断标志物有助于优化乳腺癌的治疗方案。乳头瘤病毒与 BC 病程进展/结果之间的直接相关性已得到证实。然而,目前瘤变相关基因(ARGs)在乳腺癌中的作用仍不明显。我们利用 METABRIC 数据集仔细研究和评估了 BC 与健康乳腺组织中表达不同的 ARGs。我们采用无监督共识ARGs聚类方法将患者分为不同的亚型。利用ESTIMATE算法评估免疫浸润模式。通过 LASSO 回归和单变量 COX 回归分析得出了预后基因表达模式。随后,利用卡普兰-梅耶生存曲线对这些特征进行了检验。采用 6 对新鲜组织标本(肿瘤和邻近非肿瘤),通过 qPCR 评估 7 个 ARGs 基因的表达。值得注意的是,DCN 和 FOS 在 BC 组织中没有表达,我们在随后的实验中排除了这两个基因。此外,在其余 5 个 ARGs 基因中,只有 ADH1A 的表达在统计上显示出新采集的癌症组织与邻近组织之间的显著差异。ADH1A高表达和ADH1A-sh载体分别转染MCF-7和MCF-7-AR细胞系。采用 RT-qPCR 和 Western 印迹技术检测了临床样本中 FABP4、CALML5、ADH1A、C1orf106、CIDEC、β-catenin、N-cadherin 和 Vimentin 的表达情况。为了评估细胞的迁移和侵袭潜力,研究人员采用了经孔室迁移和侵袭技术。利用细胞计数试剂盒-8(CCK-8)检测法对细胞增殖进行了详细评估。细胞凋亡指数是通过流式细胞仪分析确定的。由七个基因(即 ADH1A、DCN、CIEDC、FABP4、FOS、CALML5 和 C1orf106)组成的创新性厌氧相关特征被设计出来,用于将 BC 患者分为高风险和低风险两组。这一独特的风险评估模型是通过独特的特征方法制定的,已被证实是一个独立的预后指标。其他分析表明,不同的风险亚型在肿瘤微环境和药物敏感性方面存在差异。抑制ADH1A会增强肿瘤的迁移和侵袭能力,并降低这些与肿瘤发生相关的蛋白水平,从而强调了ADH1A在BC进展中的预后作用。通过细致的研究,我们阐明了ARGs在BC中可能的分子标记和临床意义。事实证明,我们的模型包含了七种 ARGs,能准确预测 BC 患者的生存预后。此外,对 ADH1A 的深入分子研究加深了我们对 BC 中 ARGs 的理解,为指导 BC 患者的个性化精准治疗开辟了一条新途径。
A comprehensive genome-based analysis identifies the anti-cancerous role of the anoikis-related gene ADH1A in modulating the pathogenesis of breast cancer.
Breast cancer (BC), a widespread and lethal neoplasm, is irrespective of the subtype of BC. Metastasis remains a crucial determinant for unfavorable outcome. The identification of novel diagnostic markers is instrumental in optimizing the treatment regime for BC. The direct correlation between anoikis and the progression/outcome of BC is well established. Nevertheless, the contribution of anoikis-related genes (ARGs) in BC remains obscure at present. We implemented the METABRIC dataset to scrutinize and assess differentially expressed ARGs in BC versus healthy breast tissues. An unsupervised consensus clustering approach for ARGs was employed to classify patients into diverse subtypes. ESTIMATE algorithms were utilized to assess immune infiltrative patterns. Prognostic gene expression patterns were derived from LASSO regression and univariate COX regression analysis. Subsequently, these signatures underwent examination via use of the Kaplan-Meier survival curve. 6 pairs of fresh tissue specimens (tumor and adjacent non-tumor) were employed to assess the expression of 7 ARGs genes via qPCR. Notably, DCN and FOS were not expressed in BC tissue, which had been excluded in our subsequent experiments. Also, among remaining 5 ARGs, solely the expression of ADH1A demonstrated a statistically remarkable disparity between freshly collected cancer tissues and the adjacent ones. ADH1A-overexpressed and ADH1A-sh vectors were transfected into MCF-7 and MCF-7-AR cell lines, respectively. The expression status of FABP4, CALML5, ADH1A, C1orf106, CIDEC, β-catenin, N-cadherin, and Vimentin in the clinical samples were scrutinized using RT-qPCR and western blotting techniques. Migration and invasion through transwell chambers were employed to assess the migratory and invasive potential of the cells. Detailed evaluation of cell proliferation was conducted utilizing a Cell Counting Kit-8 (CCK-8) assay. The apoptotic index of the cells was determined by flow cytometry analysis. An innovative anoikis-associated signature consisting of seven genes, namely ADH1A, DCN, CIEDC, FABP4, FOS, CALML5, and C1orf106, was devised to stratify BC patients into high- and low-risk cohorts. This unique risk assessment model, formulated via the distinctive signature approach, has been validated as an independent prognostic indicator. Additional analysis demonstrated that distinct risk subtypes manifested variances in the tumor microenvironment and drug sensitivities. Suppression of ADH1A enhanced the migratory and invasive capacities and reduced these tumorigenesis-related protein levels, underscoring the prognostic role of ADH1A in the progression of BC. Through our meticulous study, we have elucidated the possible molecular markers and clinical implications of ARGs in BC. Our model, which incorporate seven ARGs, has proven to accurately forecast the survival outcomes of BC patients. Moreover, the thorough molecular study of ADH1A has augmented our comprehension of ARGs in BC and opened a novel avenue for guiding personalized and precise therapeutic interventions for BC patients.
期刊介绍:
Molecular Genetics and Genomics (MGG) publishes peer-reviewed articles covering all areas of genetics and genomics. Any approach to the study of genes and genomes is considered, be it experimental, theoretical or synthetic. MGG publishes research on all organisms that is of broad interest to those working in the fields of genetics, genomics, biology, medicine and biotechnology.
The journal investigates a broad range of topics, including these from recent issues: mechanisms for extending longevity in a variety of organisms; screening of yeast metal homeostasis genes involved in mitochondrial functions; molecular mapping of cultivar-specific avirulence genes in the rice blast fungus and more.