Abstract：
As a promising immune checkpoint of immunogenic cell death (ICD) and multifunctional calcium-binding molecular chaperone, calreticulin (CALR) has been attracting increasing attention. CALR mainly locates in cellular endoplasmic reticulum and significantly affects cell proliferation, invasion, induction of apoptosis, and angiogenesis in breast invasive carcinoma (BRCA). CALR overexpression might be correlated with a worse outcome. Nonetheless, it remains obscure how CALR correlates with immune infiltration and survival prognosis of BRCA. In this study, we investigated CALR expression utilizing RNAseq data from the cancer genome atlas (TCGA) and genotype-tissue expression (GTEx) database. The prognostic value of CALR was analyzed using clinical survival data. Enrichment analysis was conducted using the R package \"clusterProfiler.\" We downloaded the immune cell infiltration score of TCGA samples from published articles and online databases and performed a correlation analysis between immune cell infiltration levels and CALR expression. We further assessed the association between CALR and immunomodulators. Moreover, we also evaluated the expression of CALR in 100 formalin-fixed and paraffin-embedded breast cancer and adjacent normal breast tissue specimens. Our results found that CALR was highly expressed in BRCA, and CALR expression levels differed in pathological stages, T stages, and N stages. Besides, these results suggested that CALR overexpression may have adverse effects on the progression-free interval (PFI) and disease-free interval (DFI), which may be related to tumor proliferation, invasion, and metastasis, leading to tumor deterioration. Meanwhile, immune cell infiltration analysis revealed a correlation between the expression of CALR and the number of neutrophils and dendritic cells, suggesting that CALR was highly correlated with many immunomodulators in BRCA. Our results provide potential biomarkers of CALR in BRCA. CALR may interact synergistically with other immunomodulators to regulate the immune microenvironment, which could be utilized to develop new immunotherapy drugs.
摘要：
作为免疫原性细胞死亡（ICD）和多功能钙结合分子伴侣的免疫检查点，钙网蛋白(CALR)已引起越来越多的关注。CALR主要位于细胞内质网,对细胞增殖有显著影响,入侵,诱导凋亡，乳腺浸润性癌（BRCA）的血管生成。CALR过表达可能与较差的结果相关。尽管如此，目前尚不清楚CALR与BRCA的免疫浸润和生存预后的关系。在这项研究中,我们利用来自癌症基因组图谱(TCGA)和基因型-组织表达(GTEx)数据库的RNAseq数据研究了CALR表达.使用临床生存数据分析CALR的预后价值。使用R包“clusterProfiler”进行富集分析。“我们从发表的文章和在线数据库下载了TCGA样本的免疫细胞浸润评分，并进行了免疫细胞浸润水平和CALR表达之间的相关性分析。我们进一步评估了CALR和免疫调节剂之间的关联。此外，我们还评估了100例福尔马林固定和石蜡包埋的乳腺癌和邻近的正常乳腺组织标本中CALR的表达。我们的结果发现CALR在BRCA中高表达，和CALR表达水平在病理阶段不同，T级,N个阶段。此外,这些结果表明，CALR过表达可能对无进展间隔（PFI）和无病间隔（DFI）有不利影响，这可能与肿瘤增殖有关，入侵,和转移，导致肿瘤恶化。同时,免疫细胞浸润分析显示CALR的表达与中性粒细胞和树突状细胞的数量之间存在相关性，提示CALR与BRCA中的多种免疫调节剂高度相关。我们的结果提供了BRCA中CALR的潜在生物标志物。CALR可能与其他免疫调节剂协同作用以调节免疫微环境。可用于开发新的免疫治疗药物。
