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山东大学学报 (医学版) ›› 2021, Vol. 59 ›› Issue (10): 25-31.doi: 10.6040/j.issn.1671-7554.0.2021.0230

• 基础医学 • 上一篇    下一篇

曲美替尼在卵巢癌中对PAX8的表达作用

王正阳1,夏艳2,师凯旋3,陶琨4,王小杰1,2   

  1. 1.蚌埠医学院研究生院, 安徽 蚌埠 233000;2.同济大学附属第一妇婴保健院妇科, 上海 200092;3.上海交通大学医学院附属仁济医院, 癌基因及相关基因国家重点实验室, 临床干细胞研究中心, 上海 200127;4.上海交通大学医学院附属同仁医院病理科, 上海 200050
  • 发布日期:2021-10-15
  • 通讯作者: 王小杰. E-mail:wangxiaojie2008@hotmail.com

Effects of Trametinib on PAX8 expression in ovarian cancer

WANG Zhengyang1, XIA Yan2, SHI Kaixuan3, TAO Kun4, WANG Xiaojie1,2   

  1. 1. Graduate School of Bengbu Medical College, Bengbu 233000, Anhui, China;
    2. Department of Gynecology, Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University, Shanghai 200092, China;
    3. State Key Laboratory of Oncogenes and Related Genes, Clinical Stem Cell Research Center, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China;
    4. Department of Pathology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200050, China
  • Published:2021-10-15

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摘要： 目的 探究MEK抑制剂—曲美替尼在卵巢癌中对PAX8表达的作用。 方法 应用基因富集分析(GSEA)检测卵巢癌细胞经曲美替尼处理后相关基因表达的差异,蛋白免疫印迹法、免疫荧光染色和实时荧光定量PCR检测曲美替尼对卵巢癌细胞PAX8蛋白表达和PAX8 mRNA的影响;结晶紫染色法检测曲美替尼对卵巢癌细胞增殖的影响;蛋白免疫印迹法、实时荧光定量PCR和CCK8实验检测干扰KRAS突变卵巢癌细胞ELK1基因后PAX8表达及细胞增殖的变化。 结果 GSEA结果显示,曲美替尼使卵巢癌细胞的KRAS基因和ELK1基因表达下调(P<0.01);实时荧光定量PCR结果显示,曲美替尼能抑制KRAS突变卵巢癌细胞中PAX8的转录(P<0.001);结晶紫染色法结果显示,曲美替尼处理后的KRAS突变卵巢癌细胞的增殖能力降低;蛋白免疫印迹法结果显示,曲美替尼能抑制KRAS突变卵巢癌细胞PAX8基因和ELK1基因表达(P<0.001),干扰KRAS突变卵巢癌细胞ELK1基因能抑制PAX8基因的表达(P<0.001);CCK8结果显示,干扰ELK1基因表达可抑制KRAS突变卵巢癌细胞增殖(P<0.05)。 结论 曲美替尼在KRAS突变的卵巢癌中通过ELK1抑制PAX8表达及细胞增殖。

关键词: 曲美替尼, 卵巢癌, PAX8蛋白, 细胞增殖, KRAS基因突变

Abstract: Objective To investigate the effects of Trametinib, a MEK inhibitor, on PAX8 expression in ovarian cancer. Methods The expression changes of related genes in ovarian cancer cells treated with Trametinib were detected with gene set enrichment analysis(GSEA). The effects of Trametinib on the protein and mRNA expressions of PAX8 in ovarian cancer cells were detected with Western blotting, immunofluorescence staining and real-time quantitative PCR. The effects of Trametinib on the proliferation of ovarian cancer cells was detected with crystal violet staining. The change of PAX8 expression and cell proliferation in KRAS mutant ovarian cancer cells transfected with si-ELK1-RNA were detected with Western blotting, real-time quantitative PCR and CCK8 experiment. Results GSEA showed that Trametinib inhibited KRAS and ELK1 genes in ovarian cancer cells(P<0.01). Real-time quantitative PCR showed that Trametinib inhibited PAX8 transcription in KRAS mutant ovarian cancer cells(P<0.001). Crystal violet staining showed that Trametinib reduced the proliferation of KRAS mutated ovarian cancer cells. Western blotting showed that Trametinib inhibited the expression of PAX8 and ELK1 genes in KRAS mutant ovarian cancer cells(P<0.001), and interference with ELK1 gene inhibited the expression of PAX8 gene in KRAS mutant ovarian cancer cells(P<0.001); The results of CCK8 showed that interfering with ELK1 gene expression inhibited the proliferation of KRAS mutant ovarian cancer cells(P<0.05). Conclusion Trametinib inhibits PAX8 expression and cell proliferation through ELK1 gene in KRAS mutant ovarian cancer.

Key words: Trametinib, Ovarian cancer, PAX8 protein, Cell proliferation, KRAS gene mutation

中图分类号: 

  • R737.31
[1] Spreafico A, Oza A, Clarke B, et al. Genotype-matched treatment for patients with advanced type I epithelial ovarian cancer[J]. Gynecologic Oncology, 2017, 5(144): 23-28.
[2] Jessmon P, Boulanger T, Zhou W, et al. Epidemiology and treatment patterns of epithelial ovarian cancer[J]. Expert Rev Anticancer Ther, 2017, 17(5): 427-437.
[3] Webb P, Jordan S. Epidemiology of epithelial ovarian cancer[J]. Best Pract Res Clin Obstet Gynaecol, 2017, 41: 3-14. doi: 10.1016/j.bpobgyn.2016.08.006.
[4] Bleu M, Mermet-Meillon F, Apfel V, et al. PAX8 and MECOM are interaction partners driving ovarian cancer[J]. Nature Communications, 2021, 12(1): 2442-2453.
[5] Chu Y, Zhu C, Wang Q, et al. Adipose-derived mesenchymal stem cells induced PAX8 promotes ovarian cancer cell growth by stabilizing TAZ protein[J]. J Cell Mol Med, 2021, 25(9): 4434-4443.
[6] Laury A, Perets R, Piao H, et al. A comprehensive analysis of PAX8 expression in human epithelial tumors[J]. Am J Surg Pathol, 2011, 35(6): 816-826.
[7] Nonaka D, Chiriboga L, Soslow RA. Expression of pax8 as a useful marker in distinguishing ovarian carcinomas from mammary carcinomas[J]. Am J Surg Pathol, 2008, 32(10): 1566-1571.
[8] Rodgers L, ÓhAinmhire E, Young A, et al. Loss of PAX8 in high-grade serous ovarian cancer reduces cell survival despite unique modes of action in the fallopian tube and ovarian surface epithelium[J]. Oncotarget, 2016, 7(22): 32785-32795.
[9] Palma T, Lucci V, Cristofaro T, et al. A role for PAX8 in the tumorigenic phenotype of ovarian cancer cells[J]. Bmc Cancer, 2014, 14: 292-230. doi: 10.1186/1471-2407-14-292.
[10] Adler E, Corona R, Lee J, et al. The PAX8 cistrome in epithelial ovarian cancer[J]. Oncotarget, 2017, 8(65): 108316-108332.
[11] Shi K, Yin X, Cai MC, et al. PAX8 regulon in human ovarian cancer links lineage dependency with epigenetic vulnerability to HDAC inhibitors[J]. eLife Sciences, 2019, 8: e44304. doi: 10.7554/eLife.44306.
[12] Gershenson D, Sun C, Wong K. Impact of mutational status on survival in low-grade serous carcinoma of the ovary or peritoneum[J]. Br J Cancer, 2015, 113(9): 1254-1258.
[13] Bollag G, Hirth P, Tsai J, et al. Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma[J]. Nature, 2010, 467(7315): 596-599.
[14] Flaherty K, Robert C, Hersey P, et al. Improved survival with MEK inhibition in BRAF-mutated melanoma[J]. N Engl J Med, 2012, 367(2): 107-114.
[15] 金剑英, 朱延安, 谢静静, 等. MEK抑制剂在K-RAS基因突变的结直肠癌细胞中的耐药机制[J]. 浙江医学, 2020, 42(2): 109-113. JIN Jianying, ZHU Yanan, XIE Jingjing, et al. Effect of MEK inhibitor trametinib on drug resistance in K-RAS mutated colorectal cancer cells and its mechanism[J]. Zhejiang Medical Journal, 2020, 42(2): 109-113.
[16] 陈晓艳, 龚一心, 焦顺昌. 西妥昔单抗联合曲美替尼对人结肠癌细胞增殖和凋亡的影响[J]. 解放军医学院学报, 2019, 40(6): 590-595. CHEN Xiaoyan, GONG Yixin, JIAO Shunchang. Effects of cetuximab combined with trametinib on proliferation and apoptosis of human colon cancer cells[J]. Academic Journal of Chinese PLA Medical School, 2019, 40(6): 590-595.
[17] Chi N, Epstein J. Getting your Pax straight: Pax proteins in development and disease[J]. Trends Genet, 2002, 18(1): 41-47.
[18] Hibbs K, Skubitz K, Pambuccian S, et al. Differential gene expression in ovarian carcinoma: identification of potential biomarkers[J]. Am J Pathol, 2004, 165(2): 397-414.
[19] Bleu M, Gaulis S, Rui L, et al. PAX8 activates metabolic genes via enhancer elements in Renal Cell Carcinoma[J]. Nature Communications, 2019, 10(1): 1-10.
[20] Elias K, Emori M, Westerling T, et al. Epigenetic remodeling regulates transcriptional changes between ovarian cancer and benign precursors[J]. Jci Insight, 2016, 1(13): e87988. doi: 10.1172/jci.insight.87988.
[21] Belardinelli A, Barabas M, Himmelbach M, et al. Anticipatory eye fixations reveal tool knowledge for tool interaction[J]. Exp Brain Res, 2016, 234(8): 2415-2431.
[22] Hansen R, Peters U, Babbar A, et al. The reactivity-driven biochemical mechanism of covalent KRASG12C inhibitors[J]. Nat Struct Mol Biol, 2018, 25(6): 454-462.
[23] Li X, Wang J, Zhang C, et al. Circular RNA circITGA7 inhibits colorectal cancer growth and metastasis by modulating the Ras pathway and upregulating transcription of its host gene ITGA7[J]. J Pathol, 2018, 246(2): 166-179.
[24] Gong S, Xu DS, Zhu JL, et al. Efficacy of the MEK inhibitor cobimetinib and its potential application to colorectal cancer cells[J]. Cell Physiol Biochem, 2018, 47(2): 680-693.
[25] Akatsu Y, Takahashi N, Yoshimatsu Y, et al. Fibroblast growth factor signals regulate transforming growth factorβ-induced endothelial-to-myofibroblast transition of tumor endothelial cells via Elk1[J]. Mol Oncol, 2019, 13(8): 1706-1724.
[26] Ma J, Liu X, Chen H, et al. The c-KIT-ERK1/2 signaling activated ELK1 and upregulated carcinoembryonic antigen expression to promote colorectal cancer progression[J]. Cancer Science, 2021, 112(2): 655-667.
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