Research Article siRNA-mediated mesothelin silencing for treatment of mesothelioma

IF 0.6 Q4 GENETICS & HEREDITY Genetics and Molecular Research Pub Date : 2022-01-01 DOI:10.4238/gmr18955

A. Aksoy, A. Varoğlu, E. Onalan, A. Tektemur, G. Artas

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Abstract

It is known that mesothelin (MSLN) is overexpressed in some cancers, and that it plays a role in cell growth through Wnt family member 1 protein. Malignant transformation usually occurs with disruption of autoregulation of autophagy-related genes. We examined the effect of MSLN on survival and clinicopathological features in mesothelioma cases, as well as variations in genes associated with autophagy, invasion, apoptosis-related genes after siRNA-mediated MSLN silencing transfection in a mesothelioma cell line (SPC212). MSLN expression was analyzed, immunohistochemically, in formalinfixed paraffin-embedded 60 mesothelioma cases. MSLN expression was categorized by median MSLN histoscores as low (L), high (H). The correlation between the levels of MSLN expression, clinicopathological features, and survival was determined in mesothelioma cases. The siRNA-mediated MSLN incubated SPC212 cells were transfected and compared to negative control siRNAs. mRNA levels were determined for autophagy, invasion, and apoptosis related-genes with RT-PCR, and western blotting in SPC212 cells after MSLN silencing. All of the cases were immunoreactive for MSLN expression. H-MSLN was associated with a favorable prognosis according to Kaplan-Meier survival analysis, but Cox regression analysis revealed that only stage was a significant independent factor for estimating survival. MSLN overexpression was

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sirna介导的间皮素沉默治疗间皮瘤

已知间皮素(mesothelin, MSLN)在某些癌症中过表达，并通过Wnt家族成员1蛋白在细胞生长中发挥作用。恶性转化通常伴随着自噬相关基因的自调节被破坏而发生。我们在间皮瘤细胞系(SPC212)中检测了sirna介导的MSLN沉默转染后，MSLN对间皮瘤患者生存和临床病理特征的影响，以及与自噬、侵袭、凋亡相关基因的变化。用免疫组织化学方法分析60例甲醛固定石蜡包埋的间皮瘤组织中MSLN的表达。根据中位MSLN组织评分将MSLN表达分为低(L)和高(H)。在间皮瘤病例中，确定MSLN表达水平、临床病理特征和生存率之间的相关性。转染sirna介导的MSLN培养的SPC212细胞，并与阴性对照sirna进行比较。在MSLN沉默后，采用RT-PCR和western blotting检测SPC212细胞中自噬、侵袭和凋亡相关基因的mRNA水平。所有病例对MSLN表达均有免疫反应。Kaplan-Meier生存分析显示，H-MSLN与预后良好相关，但Cox回归分析显示，只有分期是估计生存的重要独立因素。MSLN过表达

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来源期刊

Genetics and Molecular Research 生物-生化与分子生物学

CiteScore

1.00

自引率

25.00%

发文量

审稿时长

3 months

期刊介绍： Genetics and Molecular Research (GMR), maintained by the Research Foundation of Ribeirão Preto (Fundação de Pesquisas Científicas de Ribeirão Preto), publishes high quality research in genetics and molecular biology. GMR reflects the full breadth and interdisciplinary nature of this research by publishing outstanding original contributions in all areas of biology. GMR publishes human studies, as well as research on model organisms—from mice and flies, to plants and bacteria. Our emphasis is on studies of broad interest that provide significant insight into a biological process or processes. Topics include, but are not limited to gene discovery and function, population genetics, evolution, genome projects, comparative and functional genomics, molecular analysis of simple and complex genetic traits, cancer genetics, medical genetics, disease biology, agricultural genomics, developmental genetics, regulatory variation in gene expression, pharmacological genomics, evolution, gene expression, chromosome biology, and epigenetics.