黏蛋白16基因通过磷酸肌醇3-激酶/蛋白激酶B通路调节胆囊癌细胞活力和迁移

李新 田蜜 彭冰 何莉莉*

doi:10.16098/j.issn.0529-1356.2019.05.012

PDF(390 KB)

解剖学报 ›› 2019, Vol. 50 ›› Issue (5) : 613-619. DOI: 10.16098/j.issn.0529-1356.2019.05.012

肿瘤生物学

黏蛋白16基因通过磷酸肌醇3-激酶/蛋白激酶B通路调节胆囊癌细胞活力和迁移

李新¹田蜜² 彭冰¹何莉莉^3*

作者信息 +

Mucin 16 regulating activity and migration of gallbladder cancer cells through phosphoinositide 3-kinase/protein kinase B pathway

LI Xin¹ TIAN Mi² PENG Bing¹ HE Li-li ^3*

Author information +

文章历史 +

摘要

目的探讨黏蛋白16基因（MUC16）是否通过磷酸肌醇3-激酶/蛋白激酶B（PI3K/Akt）通路调节胆囊癌细胞（GBC-SD）活力和迁移。方法过表达MUC16后，通过Real-time PCR筛选相关基质金属蛋白酶（MMP）家族的蛋白质。其次，过表达MUC16、敲低MUC16和PI3K/Akt通路抑制剂BKM120处理后，通过免疫印迹法检测PI3K/Akt通路和MMP-9的蛋白水平。最后，过表达MUC16或敲低MUC16，通过细胞计数试剂盒8（CCK-8）和刮伤实验检测GBC-SD的活力和迁移情况。结果过表达MUC16后，MMP-9 mRNA的相对表达量显著升高（P<0.05）。过表达MUC16后， MMP-9、p-Akt、PI3K的蛋白水平明显升高（P<0.05），但是PI3K抑制剂BKM120可以避免这现象。而敲低MUC16后，发现MMP-9、p-Akt、PI3K的蛋白水平明显降低（P<0.05）。过表达MUC16后，GBC-SD的细胞活力和迁移能力明显增高（P<0.05），而敲低了MUC16后，GBC-SD的细胞活力和迁移能力明显降低（P<0.05）。另外，敲低MMP-9后，GBC-SD的细胞活力和迁移能力也明显降低（P<0.05）。但是，在过表达MUC16的同时敲低MMP-9，发现GBC-SD的细胞活力和迁移能力与对照组相比差异无显著性（P>0.05）。结论 MUC16激活PI3K/Akt通路促进MMP-9的蛋白表达，进而提升胆囊癌细胞GBC-SD的细胞活力和迁移能力。

Abstract

Objective To investigate the effect of mucin 16(MUC16) on the viability and migration of gallbladder cancer cells (GBC-SD) through phospho inositide 3-kinase/protein kinase B (PI3k/Akt) pathway. Methods Firstly, after overexpression of MUC16, the matrix metalloproteinase(MMP) family proteins were screened by Real-time PCR. Secondly, after overexpression of MUC16, knockdown of MUC16 and PI3K/Akt pathway inhibitor BKM120, the protein levels of PI3K/Akt pathway and MMP-9 were detected by Western blotting. Finally, the proliferation, viability and migration of gallbladder cancer cells were detected by cell counting kit-8 (CCK-8) and scratch test. Results After overexpression of MUC16, the relative expression of mRNA in MMP-9 increased significantly (P<0.05). After overexpression of MUC16, the levels of MMP-9, p-Akt and PI3K increased significantly (P<0.05), but PI3K inhibitor BKM120 could avoid this phenomenon. After knocking down MUC16, the protein levels of MMP-9, p-Akt and PI3K decreased significantly (P<0.05).After overexpression of MUC16, the cell viability and migration ability of GBC-SD increased significantly (P<0.05), while after knocking down MUC16, the cell viability and migration ability of GBC-SD decreased significantly (P<0.05). In addition, after knocking down MMP-9, the cell viability and migration ability of GBC-SD also decreased significantly (P<0.05). However, when MUC16 was overexpressed and MMP-9 was knocked down, there was no significant difference in cell viability and migration ability between GBC-SD and the control group (P>0.05). Conclusion MUC16 activates PI3K/Akt pathway to promote the expression of MMP-9 protein, thereby enhancing the cell viability and migration of GBC-SD.

导出引用

李新田蜜彭冰何莉莉. 黏蛋白16基因通过磷酸肌醇3-激酶/蛋白激酶B通路调节胆囊癌细胞活力和迁移[J]. 解剖学报. 2019, 50(5): 613-619 https://doi.org/10.16098/j.issn.0529-1356.2019.05.012

LI Xin TIAN Mi PENG Bing HE Li-li. Mucin 16 regulating activity and migration of gallbladder cancer cells through phosphoinositide 3-kinase/protein kinase B pathway[J]. Acta Anatomica Sinica. 2019, 50(5): 613-619 https://doi.org/10.16098/j.issn.0529-1356.2019.05.012

参考文献

［1］ Gao PY. Clinicopathological features of primary gallbladder carcinoma［J］. Chinese Continuing Medical Education, 2017,9(22):74-76. (in Chinese)

高萍宇. 原发性胆囊癌的临床病理特点［J］. 中国继续医学教育, 2017，9(22):74-76.

［2］ Li Q. The challenge of surgical treatment of gallbladder carcinoma［J］. Journal of Clinical Surgery, 2017, 25(6): 405-407. (in Chinese)

李强. 胆囊癌外科治疗的挑战［J］. 临床外科杂志, 2017, 25(6):405-407.

［3］ Li ZhJ, Wang LX, Li YK, et al. Clinicopathological analysis of mucinous inflammatory fibroblastic sarcoma［J］. Hainan Medical Journal, 2017,28(24): 4079-4080. (in Chinese)

李正金, 王丽霞, 李耀康,等. 黏液炎症性纤维母细胞肉瘤临床病理分析［J］. 海南医学, 2017，28(24):4079-4080.

［4］ Chen X, Wang XP. Research progress of MUC16 in the development of ovarian cancer［J］. International Journal of Obstetrics and Gynecology, 2016, 43(5): 506-509. (in Chinese)

陈昕, 汪希鹏. MUC16在卵巢癌发生发展中的研究进展［J］. 国际妇产科学杂志, 2016, 43(5):506-509.

［5］ Th ériault C,Pinard M,Comamala M,et al.MUC16(CA125)regulates epithelial ovarian cancer cell growth,tumorigenesis and metastasis［J］.Gynecol Oncol,2011,121(3):434-443.

［6］ He X, Zhang MH, Lu Y, et al. Protection and regulation mechanism of mucin in intestinal mucosal barrier［J］. Medical Review, 2017, 23(1):1-5. (in Chinese)

贺欣, 张目涵, 路瑶,等. 黏蛋白在肠黏膜屏障中的保护调节机制［J］. 医学综述, 2017, 23(1):1-5.

［7］ Wang K, Xu Q, Zuo YZh, et al. Inhibitory effect of dihydromyricetin combined with etoposide on JAR of choriocarcinoma cells ［J］. Acta Anatomica Sinica, 2018,49(3): 330-336. (in Chinese)

王康,许倩,左彦珍,等.二氢杨梅素联合依托泊苷对绒毛膜癌细胞JAR的抑制作用［J］.解剖学报,2018,49(3):330-336.

［8］ Li DD, Cui ShY, Jia LM, et al. Advances in the formation of pre-metastatic microenvironment in tumor metastasis［J］. Modern Oncology Medicine, 2018,26(6): 962-965. (in Chinese)

李丹丹, 崔守烨, 贾立敏,等. 肿瘤转移中预转移微环境形成的研究进展［J］. 现代肿瘤医学, 2018，26(6): 962-965.

［9］ Das S,Batra SK.Understanding the unique attributes of MUC16(CA125):potential implications in targeted therapy［J］.Cancer Res,2015,75(22):4669-4674.

［10］Bressan A,Bozzo F,Maggi CA,et al.OC125,M11 and OV197 epitopes are not uniformly distributed in the tandemrepeat region of CA125 and require the entire SEA domain［J］.Dis Markers,2013,34(4):257-267.

［11］Ricardo S,Marcos-Silva L,Pereira D,et al.Detection of glycomucin profiles improves specificity of MUC16 and MUC1 biomarkers in ovarian serous tumours［J］.Mol Oncol,2015,9(2):503-512.

［12］Chugh S,Gnanapragassam VS,Jain M,et al.Pathobiological implications of mucin glycans in cancer:Sweet poison and novel targets［J］.Biochim Biophys Acta,2015,1856(2):211-225.

［13］Nemunaitis JM, Brownglabeman U, Soares H, et al. Gallbladder cancer: review of a rare orphan gastrointestinal cancer with a focus on populations of New Mexico［J］. BMC Cancer, 2018, 18(1):665.

［14］Pastan I,Hassan R.Discovery of mesothelin and exploiting it as a target for immunotherapy［J］. Cancer Res,2014,74(11):2907-2912.

［15］Vinay DS,Ryan EP,Pawelec G,et al.Immune evasion in cancer:Mechanistic basis and therapeutic strategies［J］.Semin Cancer Biol,2015,35(Suppl):S185-S198.

［16］Madsen CB,Lavrsen K,Steentoft C,et al.Glycan elongation beyond the mucin associated Tn antigen protects tumor cells from immune mediated killing［J］.PLoS One,2013,8(9):e72413.

［17］Coelho R, Marcossilva L, Ricardo S, et al. Peritoneal dissemination of ovarian cancer: role of MUC16mesothelin interaction and implications for treatment.［J］. Expert Rev Anticancer Ther, 2018, 18(2):177-186.

［18］Comamala M,Pinard M,Thériault C,et al.Downregulation of cell surface CA125/MUC16 induces epithelialtomesenchymal transition and restores EGFR signalling in NIH:OVCAR3 ovarian carcinoma cells［J］.Br J Cancer,2011,104(6):989-999.

［19］Cheng X, Yang Y, Fan Z, et al. MKL1 potentiates lung cancer cell migration and invasion by epigenetically activating MMP9 transcription［J］. Oncogene, 2015, 34(44):5570-5581.

［20］Li BSh, Hong L, Cheng LW, et al. Expression of phosphatidylinositol 3-kinase-protein kinase B-forkhead box protein signaling pathway in pelvic organ prolapse［J］. Chinese Journal of Family Planning and Obstetrics and Gynecology, 2017, 9 (4): 27-29. (in Chinese)

李秉枢, 洪莉, 程丽薇,等. 磷脂酰肌醇3激酶-蛋白激酶B-叉头框蛋白信号通路在盆腔脏器脱垂中的表达［J］. 中国计划生育和妇产科, 2017, 9(4):27-29.

［21］Das S,Rachagani S,Torres-Gonzalez MP,et al.Carboxyl-terminal domain of MUC16 imparts tumorigenic and metastatic functions through nuclear translocation of JAK2 to pancreatic cancer cells［J］.Oncotarget,2015,6(8):5772-5787.

［22］Giannakouros P,Comamala M,Matte Ⅰ,et al.MUC16 mucin(CA125)regulates the formation of multicellular aggregates by altering β-catenin signaling［J］.Am J Cancer Res,2014,5(1):219-230.

［23］Rao TD,Tian H,Ma X,et al.Expression of the carboxy-terminal portion of MUC16/CA125 induces transformation and tumor invasion［J］.PLoS One,2015,10(5):e0126633.

［24］Chen SH, Hung WC, Wang P, et al. Mesothelin Binding to CA125/MUC16 promotes pancreatic cancer cell motility and invasion via MMP-7 activation［J］. Scientific Reports, 2013, 3(5):1870.

［25］Lakshmanan Ⅰ, Ponnusamy MP, Das S, et al. MUC16 induced rapid G2/M transition via interactions with JAK2 for increased proliferation and anti-apoptosis in breast cancer cells［J］. Oncogene, 2012, 31(7):805-817.

［26］Liu Q, Cheng Z, Luo L, et al. C-terminus of MUC16 activates Wnt signaling pathway through its interaction with β-catenin to promote tumorigenesis and metastasis［J］. Oncotarget, 2016, 7(24):36800-36813.