Wei Li, Chang Haiyan, Huo Song
Department of Pathology, Jining First People's Hospital, Jining, Shandong 272111, P.R. China.
Department of Gastrointestinal Surgery, Jining First People's Hospital, Jining, Shandong 272111, P.R. China.
Oncol Lett. 2019 Feb;17(2):1807-1811. doi: 10.3892/ol.2018.9750. Epub 2018 Nov 22.
Mutations of K-rat sarcoma (K-ras) in patients with cardia cancer and their effects on the expression of fascin were investigated. A total of 90 cardia cancer patients treated in Jining First People's Hospital from March 2014 to March 2017 were randomly selected. Genomic deoxyribonucleic acid (DNA) was extracted from paraffin-embedded cardia cancer specimens. Pyrosequencing was applied to detect sequences of K-ras gene in all patients and to analyze the mutations of K-ras gene. Then, genotyping of mutations at each mutation site was carried out using quantitative polymerase chain reaction (qPCR). The expression level of fascin in patients was measured via immunohistochemistry and qPCR. The results revealed that among 90 patients with cardia cancer, 21 patients had K-ras mutations (23.3%), including 20 cases of exon 12 mutation and 1 case of exon 13 mutation. Risk factor analyses revealed that alcohol abuse was a high risk factor for mutations (p<0.05). There was no significant difference in the mutation probability between heterozygotes and homozygotes for four mutations at codon 12 (p>0.05). The heterozygote at codon 13 had a higher mutation probability than homozygote (p<0.05). Immunohistochemistry suggested that the number of positive cells in the mutant group was larger than that in the non-mutant group (p<0.05). The results of qPCR showed that the expression level of fascin gene in the mutant group was 2.3 times higher than that in the non-mutant group (p<0.05). In conclusion, the probability of codon 12 mutation in K-ras gene is increased in patients with cardia cancer, and fascin is highly expressed in mutant patients, which is positively correlated with the mutations in K-ras gene.
研究了贲门癌患者K-鼠肉瘤(K-ras)突变及其对成束蛋白表达的影响。随机选取2014年3月至2017年3月在济宁市第一人民医院接受治疗的90例贲门癌患者。从石蜡包埋的贲门癌标本中提取基因组脱氧核糖核酸(DNA)。采用焦磷酸测序法检测所有患者的K-ras基因序列,并分析K-ras基因的突变情况。然后,使用定量聚合酶链反应(qPCR)对每个突变位点的突变进行基因分型。通过免疫组织化学和qPCR检测患者中成束蛋白的表达水平。结果显示,90例贲门癌患者中,21例发生K-ras突变(23.3%),其中外显子12突变20例,外显子13突变1例。危险因素分析显示,酗酒是突变的高危因素(p<0.05)。密码子12处四种突变的杂合子和纯合子之间的突变概率无显著差异(p>0.05)。密码子13处的杂合子比纯合子具有更高的突变概率(p<0.05)。免疫组织化学显示,突变组阳性细胞数多于非突变组(p<0.05)。qPCR结果显示,突变组中成束蛋白基因的表达水平是非突变组的2.3倍(p<0.05)。综上所述,贲门癌患者K-ras基因密码子12突变概率增加,成束蛋白在突变患者中高表达,且与K-ras基因突变呈正相关。