Abstract：
BACKGROUND: Breast cancer (BC) is the world\'s largest tumor species in which hormone receptor-positive patients have relatively good prognosis. However, majority of patients will develop late resistance, one of the important factors is due to the loss of the original estrogen receptor (ER) expression.
METHODS: We conducted this study in 115 patients with BC who experienced second biopsy at Jiangsu Province Hospital (JSPH) and divided patients into two subgroups ER + to - and ER + to + . First, clinicopathological characteristics between two groups were evaluated. Second, we explored candidate genes related to BC ER intratumor heterogeneity by applying next-generation sequencing (NGS) in 42 patients. Multi-omics integrative analysis of tumor transcriptomic, cancer-related pathway, diagnostic and prognostic value and immune profile were conducted. Besides, preliminary assay were also used to evaluate the correlation between KMT2C and ERα (ESR1) expression. The CCK-8, 5-Ethynyl-2\'-deoxyuridine (EdU) assays, Transwell assays and the wound scratch tests were applied to explore the cellular interactions between KMT2C and BC.
RESULTS: We find the histological type (p = 0.008) and disease-free survival (DFS) (p = 0.004) were significantly different in two subgroups. In Cox survival analysis, metastasis (Hazard ratio (HR) > 1, p = 0.007) and neo-adjuvant (HR < 1, p < 0.001) are independent prognostic factors of DFS. Besides, by analyzing NGS results, we found four genes KMT2C, FGFR19, FGF1 and FGF4 were highly mutated genes in ER + to - subgroup. Furthermore, the gene KMT2C displayed significant diagnostic value and prognostic value in BC and pan-cancer. In addition, a positive correlation between KMT2C expression and immune infiltrating levels of T cell CD4 + , macrophage and neutrophil was found. In the end, Western blot and RT-qPCR assay were used and found KMT2C and ERα (ESR1) expressions are strongly positive correlated in mRNA and protein level. Inhibition of KMT2C significantly reduced proliferation, invasion, and migration of MCF7 cells.
CONCLUSIONS: People in two cohorts from JSPH presented different clinical characteristics and prognosis. The gene KMT2C may affect the progression of BC by regulating the molecular, epigenetic activity and immune infiltration. It may also serve as a novel prognostic biomarker for BC patients who underwent ER status converted from positive to negative.
METHODS: We conducted this study in 115 patients with BC who experienced second biopsy at Jiangsu Province Hospital (JSPH) and divided patients into two subgroups ER + to - and ER + to + . First, clinicopathological characteristics between two groups were evaluated. Second, we explored candidate genes related to BC ER intratumor heterogeneity by applying next-generation sequencing (NGS) in 42 patients. Multi-omics integrative analysis of tumor transcriptomic, cancer-related pathway, diagnostic and prognostic value and immune profile were conducted. Besides, preliminary assay were also used to evaluate the correlation between KMT2C and ERα (ESR1) expression. The CCK-8, 5-Ethynyl-2\'-deoxyuridine (EdU) assays, Transwell assays and the wound scratch tests were applied to explore the cellular interactions between KMT2C and BC.
RESULTS: We find the histological type (p = 0.008) and disease-free survival (DFS) (p = 0.004) were significantly different in two subgroups. In Cox survival analysis, metastasis (Hazard ratio (HR) > 1, p = 0.007) and neo-adjuvant (HR < 1, p < 0.001) are independent prognostic factors of DFS. Besides, by analyzing NGS results, we found four genes KMT2C, FGFR19, FGF1 and FGF4 were highly mutated genes in ER + to - subgroup. Furthermore, the gene KMT2C displayed significant diagnostic value and prognostic value in BC and pan-cancer. In addition, a positive correlation between KMT2C expression and immune infiltrating levels of T cell CD4 + , macrophage and neutrophil was found. In the end, Western blot and RT-qPCR assay were used and found KMT2C and ERα (ESR1) expressions are strongly positive correlated in mRNA and protein level. Inhibition of KMT2C significantly reduced proliferation, invasion, and migration of MCF7 cells.
CONCLUSIONS: People in two cohorts from JSPH presented different clinical characteristics and prognosis. The gene KMT2C may affect the progression of BC by regulating the molecular, epigenetic activity and immune infiltration. It may also serve as a novel prognostic biomarker for BC patients who underwent ER status converted from positive to negative.
摘要：
背景：乳腺癌是世界上最大的肿瘤，激素受体阳性患者预后相对较好。然而,大多数患者会出现晚期耐药性，重要因素之一是由于原始雌激素受体（ER）表达的丧失。
方法：我们在江苏省人民医院（JSPH）进行了第二次活检的115例BC患者中进行了这项研究，并将患者分为两个亚组ERto-和ERto-。首先,评估两组患者的临床病理特征。第二,我们在42例患者中应用下一代测序(NGS),探索了与BCER瘤内异质性相关的候选基因.肿瘤转录组学的多组学整合分析，癌症相关途径，进行了诊断和预后价值以及免疫概况。此外,初步实验还用于评估KMT2C和ERα（ESR1）表达之间的相关性。CCK-8，5-乙炔基-2'-脱氧尿苷（EdU）测定，应用Transwell测定和伤口划痕测试来探索KMT2C和BC之间的细胞相互作用。
结果：我们发现两个亚组的组织学类型（p=0.008）和无病生存期（DFS）（p=0.004）显着不同。在Cox生存分析中，转移（危险比（HR）>1，p=0.007）和新辅助（HR<1，p<0.001）是DFS的独立预后因素。此外,通过分析NGS结果，我们发现了四个基因KMT2C,FGFR19，FGF1和FGF4是ER至-亚组中高度突变的基因。此外，KMT2C基因在BC和泛癌症中显示出显著的诊断价值和预后价值.此外,KMT2C表达与T细胞CD4+免疫浸润水平呈正相关,发现巨噬细胞和中性粒细胞。最后,Westernblot和RT-qPCR检测发现，KMT2C和ERα（ESR1）在mRNA和蛋白水平的表达呈极显著正相关。抑制KMT2C显著降低增殖，入侵,和MCF7细胞的迁移。
结论：来自JSPH的两组患者具有不同的临床特征和预后。KMT2C基因可能通过分子调控影响BC的进展,表观遗传活性和免疫浸润。它还可以作为接受ER状态从阳性转变为阴性的BC患者的新型预后生物标志物。
方法：我们在江苏省人民医院（JSPH）进行了第二次活检的115例BC患者中进行了这项研究，并将患者分为两个亚组ERto-和ERto-。首先,评估两组患者的临床病理特征。第二,我们在42例患者中应用下一代测序(NGS),探索了与BCER瘤内异质性相关的候选基因.肿瘤转录组学的多组学整合分析，癌症相关途径，进行了诊断和预后价值以及免疫概况。此外,初步实验还用于评估KMT2C和ERα（ESR1）表达之间的相关性。CCK-8，5-乙炔基-2'-脱氧尿苷（EdU）测定，应用Transwell测定和伤口划痕测试来探索KMT2C和BC之间的细胞相互作用。
结果：我们发现两个亚组的组织学类型（p=0.008）和无病生存期（DFS）（p=0.004）显着不同。在Cox生存分析中，转移（危险比（HR）>1，p=0.007）和新辅助（HR<1，p<0.001）是DFS的独立预后因素。此外,通过分析NGS结果，我们发现了四个基因KMT2C,FGFR19，FGF1和FGF4是ER至-亚组中高度突变的基因。此外，KMT2C基因在BC和泛癌症中显示出显著的诊断价值和预后价值.此外,KMT2C表达与T细胞CD4+免疫浸润水平呈正相关,发现巨噬细胞和中性粒细胞。最后,Westernblot和RT-qPCR检测发现，KMT2C和ERα（ESR1）在mRNA和蛋白水平的表达呈极显著正相关。抑制KMT2C显著降低增殖，入侵,和MCF7细胞的迁移。
结论：来自JSPH的两组患者具有不同的临床特征和预后。KMT2C基因可能通过分子调控影响BC的进展,表观遗传活性和免疫浸润。它还可以作为接受ER状态从阳性转变为阴性的BC患者的新型预后生物标志物。
