Jeeyar Vinay, Besra Kusumbati, Singh Shivaram Prasad, Dixit Manjusha
School of Biological Sciences, National Institute of Science Education and Research, Bhubaneswar, 752050, Odisha, India.
Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai, 400094, India.
Sci Rep. 2025 Jul 31;15(1):27958. doi: 10.1038/s41598-025-13833-3.
Gallbladder cancer (GBC) poses a significant global health threat, claiming 89,031 lives worldwide in 2022. This malignancy exhibits distinct genetic susceptibility patterns among populations, coupled with its aggressive nature and dismal prognosis. Among the factors implicated in cancer progression, Matrix Metalloproteinase 7 (MMP7) has emerged as a crucial biomarker due to its dual role in cancer causation and progression, but its role in GBC is completely unexplored. We hypothesized that promoter SNPs could modulate MMP7 expression, thus impacting GBC susceptibility. Our study aimed to assess the aberrant expression levels and genetic polymorphisms of MMP7 as potential risk factors for GBC. Initial screening of MMP7 promoter polymorphisms in a cohort of fifty GBC patients and controls using Sanger sequencing was followed by validation in a larger sample set comprising 300 GBC cases and 300 controls. Additionally, MMP7 protein expression was examined through immunohistochemistry in GBC tissue samples. Functional analysis of identified risk variants was carried out using luciferase reporter assays and in vitro promoter analysis in two cell lines to elucidate their impact on gene expression. MMP7 promoter variants, rs113823671 A > C (p-value = 0.034) and rs17098318 G > A (p-value = 0.024), exhibited significant associations with GBC. Reporter assays in two cell lines indicated that carriers of the risk allele 'A' at rs113823671 exhibited increased luciferase reporter activity, whereas no significant reporter activity was observed at rs17098318. MMP7 expression levels were markedly elevated in GBC compared to adjacent uninvolved normal tissue. Furthermore, genotype-phenotype analysis in tissue samples revealed that risk allele carriers exhibited higher MMP7 expression levels, with the correlation aligning with genetic association models. In conclusion, our study unveils the association of two novel MMP7 promoter risk variants, rs113823671, and rs17098318, with GBC and their role in GBC pathogenesis.
胆囊癌（GBC）对全球健康构成重大威胁，2022年全球有89,031人死于该病。这种恶性肿瘤在不同人群中表现出独特的遗传易感性模式，且具有侵袭性和预后不良的特点。在与癌症进展相关的因素中，基质金属蛋白酶7（MMP7）因其在癌症发生和进展中的双重作用而成为一种关键生物标志物，但其在GBC中的作用尚未得到研究。我们假设启动子单核苷酸多态性（SNP）可调节MMP7表达，从而影响GBC易感性。我们的研究旨在评估MMP7的异常表达水平和基因多态性作为GBC的潜在危险因素。首先使用桑格测序法在50例GBC患者和对照人群中对MMP7启动子多态性进行初步筛查，随后在一个更大的样本集（包括300例GBC病例和300例对照）中进行验证。此外，通过免疫组织化学检测GBC组织样本中的MMP7蛋白表达。使用荧光素酶报告基因检测和在两种细胞系中进行体外启动子分析对鉴定出的风险变异进行功能分析，以阐明它们对基因表达的影响。MMP7启动子变异rs113823671 A＞C（p值 = 0.034）和rs17098318 G＞A（p值 = 0.024）与GBC显著相关。在两种细胞系中的报告基因检测表明，rs113823671风险等位基因“A”的携带者荧光素酶报告基因活性增加，而rs17098318未观察到显著的报告基因活性。与相邻未受累的正常组织相比，GBC中MMP7表达水平明显升高。此外，组织样本中的基因型-表型分析表明，风险等位基因携带者的MMP7表达水平较高，这种相关性与遗传关联模型一致。总之，我们的研究揭示了两种新的MMP7启动子风险变异rs113823671和rs17098318与GBC的关联及其在GBC发病机制中的作用。
