YKL-40对卵巢癌SKOV-3细胞迁移能力的影响

doi:10.6040/j.issn.1671-7554.0.2016.1304

摘要/Abstract

摘要： 目的探讨YKL-40对卵巢癌SKOV-3细胞体外迁移能力的影响。方法采用RT-PCR检测卵巢癌SKOV-3细胞YKL-40 mRNA水平的表达;采用慢病毒感染的方法在SKOV-3细胞过表达YKL-40,qRT-PCR、ELISA、Western blotting验证YKL-40的过表达;Transwell实验检测YKL-40过表达对SKOV-3细胞迁移能力的影响;特异性抑制剂抑制YKL-40过表达SKOV-3细胞中的p38 MAPK通路,检测其对细胞迁移能力的影响。结果 SKOV-3细胞中未检测到YKL-40 mRNA的表达;成功构建了YKL-40过表达细胞株LV-SKOV-3,感染率达90%以上;与空载体慢病毒感染的对照SKOV-3细胞(NC-SKOV-3)相比,LV-SKOV-3细胞YKL-40无论mRNA表达还是蛋白水平(细胞总蛋白、分泌蛋白)均明显上调;Transwell细胞迁移实验表明过表达YKL-40的LV-SKOV-3细胞较空载体对照细胞NC-SKOV-3迁移能力显著增强,而通过SB203580抑制p38 MAPK通路p38蛋白磷酸化后,LV-SKOV-3细胞的迁移能力明显下调。结论 YKL-40可显著促进卵巢癌SKOV-3细胞的迁移,这一作用可能是通过p38 MAPK通路介导的。

关键词: 卵巢癌, 细胞迁移, p38 MAPK通路, YKL-40

Abstract: Objective To investigate the effect of YKL-40 on epithelial ovarian cancer SKOV-3 cell migration in vitro and to explore the underlying mechanism. Methods YKL-40 mRNA expression in SKOV-3 cells was detected by RT-PCR. SKOV-3 cells were transfected with modified lentiviral vectors containing YKL-40 gene and YKL-40 overexpression was validated by qRT-PCR, ELISA and Western blotting. Transwell assay was used to evaluate the effect of YKL-40 on SKOV-3 cell migration. p38MAPK pathway was repressed by specific inhibitors and its effect on YKL-40 overexpressed SKOV-3 cell migration was further investigated. Results YKL-40 mRNA was not detected in SKOV-3 cells. After lentivirus transfection and puromycin selection, YKL-40 overexpressed cell line LV-SKOV-3 and negative control cell line NC-SKOV-3 were established. Further analyses confirmed YKL-40 was significantly increased in LV-SKOV-3 cells on both mRNA and protein levels compared to NC-SKOV-3. After YKL-40 overexpression, the migration ability of LV-SKOV-3 was enhanced while LV-SKOV-3 cell migration was decreased after inhibition of p38MAPK pathway via SB203580. Conclusion YKL-40 significantly promotes the migration ability of SKOV-3 cells, which is partly mediated by p38MAPK pathway.

Key words: Cell migration, p38MAPK pathway, Ovarian cancer, YKL-40

中图分类号:

R737.31

陆衡,刘延国,李曙光,陈晓康,田琦,衣翠华,王秀问. YKL-40对卵巢癌SKOV-3细胞迁移能力的影响[J]. 山东大学学报（医学版）, 2017, 55(1): 33-38.

LU Heng, LIU Yanguo, LI Shuguang, CHEN Xiaokang, TIAN Qi, YI Cuihua, WANG Xiuwen. Regulation of epithelial ovarian carcinoma SKOV-3 cell migration by YKL-40[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(1): 33-38.

参考文献

[1] 李燕, 尉蔚, 吕树卿, 等. PARP-1对卵巢癌血管生成的影响[J].山东大学学报(医学版), 2014, 52(4):97-106. LI Yan, WEI Wei, L(¨overU)Shuqing, et al. Effects of PARP-1 on angiogenesis of ovarian cancer[J]. Journal of Shandong University(Health Science), 2014, 52(4):97-106.
[2] Lengyel E. Ovarian cancer development and metastasis[J]. Am J Pathol, 2010, 177(3):1053-1064.
[3] Johansen JS, Jensen BV, Roslind A, et al. Serum YKL-40, a new prognostic biomarker in cancer patients?[J]. Cancer Epidemiol Biomarkers Prev, 2006, 15(2):194-202.
[4] Jefri M, Huang YN, Huang WC, et al. YKL-40 regulated epithelial-mesenchymal transition and migration/invasion enhancement in non-small cell lung cancer[J]. BMC Cancer, 2015, 15:590. doi:10.1186/s12885-015-1592-3.
[5] Thongsom S, Chaocharoen W, Silsirivanit A, et al. YKL-40/chitinase-3-like protein 1 is associated with poor prognosis and promotes cell growth and migration of cholangiocarcinoma[J]. Tumour Biol, 2016, 37(7):9451-9463.
[6] Koul HK, Pal M, Koul S. Role of p38 MAP kinase signal transduction in solid tumors[J]. Genes & Cancer, 2013, 4(9-10):342-359.
[7] Jelovac D, Armstrong DK. Recent progress in the diagnosis and treatment of ovarian cancer[J]. CA Cancer J Clin, 2011, 61(3):183-203.
[8] Ohno M, Bauer PO, Kida Y, et al. Quantitative real-time PCR analysis of YKL-40 and its comparison with mammalian chitinase mRNAs in normal human tissues using a single standard DNA [J]. Int J Mol Sci, 2015, 16(5):9922-9935.
[9] Ober C, Chupp GL. The chitinase and chitinase-like proteins: a review of genetic and functional studies in asthma and immune-mediated diseases[J]. Curr Opin Allergy Clin Immunol, 2009, 9(5):401-408.
[10] Kawada M, Seno H, Kanda K, et al. Chitinase 3-like 1 promotes macrophage recruitment and angiogenesis in colorectal cancer[J]. Oncogene, 2012, 31(26):3111-3123.
[11] Zou L, He X, Zhang JW. The efficacy of YKL-40 and CA125 as biomarkers for epithelial ovarian cancer[J]. Braz J Med Biol Res, 2010, 43(12):1232-1238.
[12] Kzhyshkowska J, Yin S, Liu T, et al. Role of chitinase-like proteins in cancer[J]. Biol Chem, 2016, 397(3):231-247.
[13] Boisen MK, Madsen CV, Dehlendorff C, et al. The prognostic value of plasma YKL-40 in patients with chemotherapy-resistant ovarian cancer treated with Bevacizumab[J]. Int J Gynecol Cancer, 2016, 26(8):1390-1398.
[14] Hogdall EV, Ringsholt M, Hogdall CK, et al. YKL-40 tissue expression and plasma levels in patients with ovarian cancer[J]. BMC Cancer, 2009, 9:8. doi:10.1186/1471-2407-9-8.
[15] Low D, Subramaniam R, Lin L, et al. Chitinase 3-like 1 induces survival and proliferation of intestinal epithelial cells during chronic inflammation and colitis-associated cancer by regulating S100A9 [J]. Oncotarget, 2015, 6(34):36535-36550.
[16] Chiang YC, Lin HW, Chang CF, et al. Overexpression of CHI3L1 is associated with chemoresistance and poor outcome of epithelial ovarian carcinoma [J]. Oncotarget, 2015, 6(37):39740-39755.
[17] Abd El-Fattah AA, Sadik NA, Shaker OG, et al. Are SMAD7 rs4939827 and CHI3L1 rs4950928 polymorphisms associated with colorectal cancer in Egyptian patients? [J]. Tumour Biol, 2016, 37(7):9387-9397.
[18] Ku BM, Lee YK, Ryu J, et al. CHI3L1(YKL-40)is expressed in human gliomas and regulates the invasion, growth and survival of glioma cells [J]. Int J Cancer, 2011,128(6):1316-1326.
[19] Li LL, Fan JT, Li DH, et al. Effects of a small interfering RNA targeting YKL-40 gene on the proliferation and invasion of endometrial cancer HEC-1A cells[J]. Int J Gynecol Cancer, 2016, 26(7):1190-1195.
[20] Hsia TC, Yu CC, Hsiao YT, et al. Cantharidin impairs cell migration and invasion of human lung cancer NCI-H460 cells via UPA and MAPK signaling pathways[J]. Anticancer Res, 2016, 36(11):5989-5997.
[21] Cheng HH, Chu LY, Chiang LY, et al. Inhibition of cancer cell epithelial mesenchymal transition by normal fibroblasts via production of 5-methoxytryptophan[J]. Oncotarget, 2016, 7(21):31243-31256.
[22] Wang J, Cui L, Feng L, et al. Isoalantolactone inhibits the migration and invasion of human breast cancer MDA-MB-231 cells via suppression of the p38 MAPK/NF-kappaB signaling pathway[J]. Oncology Reports, 2016, 36(3):1269-1276.
[23] Zhao W, Lu M, Zhang Q. Chloride intracellular channel 1 regulates migration and invasion in gastric cancer by triggering the ROS-mediated p38 MAPK signaling [J]. Mol Med Rep, 2015, 12(6):8041-8047.

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