MAPK13调控胆管癌细胞增殖、侵袭和上皮间质转化进程的机制研究

目的 本研究旨在分析丝裂原活化蛋白激酶13(MAPK13)在胆管癌组织及细胞中的表达水平并探讨其在胆管癌细胞增殖、侵袭和上皮间质转化(EMT)中的作用。方法 通过微阵列分析筛选胆管癌组织中差异表达的mRNA，通过实时荧光定量PCR(qRT-PCR)、蛋白印迹及免疫组化染色验证MAPK13在肿瘤组织和细胞中的表达情况。进一步通过细胞转染构建MAPK13敲低(si-MAPK13)后，通过细胞活力实验检测胆管癌细胞增殖能力，流式细胞仪检测细胞凋亡，Transwell法检测细胞的迁移和侵袭性。最后，通过裸鼠成瘤实验检测MAPK13对体内成瘤及相关蛋白表达的影响。结果 本研究通过微阵列分析发现MAPK13在肿瘤组织中的表达存在显著差异，进一步通过qRT-PCR、蛋白印迹和免疫组化染色分析发现，与癌旁组织比较，MAPK13在mRNA和蛋白质水平的表达情况在肿瘤组织及细胞中均显著升高且差异具有统计学意义(t=27.92，6.90；均P<0.01)。进一步细胞活力检测表明，抑制MAPK13的表达后细胞增殖数量明显减少(F=32.13，P<0.01)。Transwell实验则显示细胞迁移和侵...

Mechanism of MAPK13 regulating the cell proliferation,invasion and epithelial-mesenchymal transition in cholangiocarcinoma

Objective The purpose of this study is to detect the expression level of mitogen activated protein kinase 13 (MAPK13) in tissues and cells of cholangiocarcinoma, and to explore its role in the cell proliferation and invasion and epithelial-mesenchymal transition (EMT) of cholangiocarcinoma. Methods The differentially expressed mRNAs in cholangiocarcinoma tissues were screened by microarray analysis, and the expressions of MAPK13 in tumor tissues and cells were verified by real-time quantitative PCR (qRT-PCR), Western blotting and immunohistochemical staining. After further constructing MAPK13 knockdown (si-MAPK13) by cell transfection, the proliferation ability of cholangiocarcinoma cells was detected by cell viability assay, cell apoptosis was detected by flow cytometry, and cell migration and invasion were detected by Transwell method. Finally, the effect of MAPK13 on tumorigenesis and the expression of related proteins in vivo were detected by nude mouse tumorigenesis experiment. Results Microarray analysis found that there were significant differences in the expression of MAPK13 in tumor tissues, and further qRT-PCR, Western blotting and immunohistochemical staining showed that, compared with adjacent tissues and normal cells, MAPK13 expressions in mRNA and protein levels were significantly increased (F=27.92, 6.90; all P<0.01). Cell viability assay showed that the number of cell proliferation was significantly reduced after inhibiting the expression of MAPK13 (F=32.13, P<0.01). Transwell assay showed that cell migration and invasion were significantly decreased (F=52.88, 38.14; all P<0.05). Compared with the control group, the expression of E-cadherin in the si-MAPK13 group was higher, while the expressions of N-cadherin and Vimentin were lower (F=77.00, 78.73, 25.9; all P<0.01). Finally, mouse tumorigenesis experiment showed that, when the expression of MAPK13 was inhibited, the tumor weight and tumor size were both significantly lower than those in the control group (F=8.994, 73.81; all P<0.01). Conclusion This study suggests that MAPK13 may regulate the progression of cholangiocarcinoma by promoting tumor cell proliferation, cell migration and invasion, and EMT.