Role of ZAC gene in the pathway of octreotide inhibiting proliferation of gastric cancer cells in vitro

doi:10.3969/j.issn.0529-1356.2013.04.009

Abstract

Abstract:

Objective To explore the role of ZAC gene in the pathway of somatostatin analogue, octreotide (OCT), inhibiting proliferation of gastric cancer cells. Methods The gastric cancer cells BGC823 and SGC7901 were treated with OCT at various concentrations for various times, respectively. MTT assay was used to screen effective condition of OCT for its antiproliferative action. After treatment with OCT (effective concentration/various times, effective time/various concentrations), the inducing effect of OCT on ZAC gene expression in gastric cancer cells was detected by Western blotting. Three fragments for ZAC gene RNA interference were inserted into pSUPER-EGFP-I vector to construct ZAC-shRNA expression vectors (pSUPER-EGFP-ZAC/1, pSUPER-EGFP-ZAC/2 and pSUPER-EGFP-ZAC/3), respectively. After identification by Hind Ⅲ/EcoRⅠdigestion and sequencing, the three ZAC-shRNA expression vectors were transfected into gastric cancer cells BGC823 and SGC7901, respectively. After G418 screening and RT-PCR identification, the gastric cancer cell lines in which ZAC gene was knock-down were established. Gastric cancer cells (control group) and those in which ZAC gene was knock-down (experimental group) were incubated with OCT at effective condition, the antiproliferative action of OCT was detected by MTT assay. Results The effective condition of OCT inhibiting proliferation of gastric cancer cells was 10nmol/L and treatmented for 24 hours. OCT induced ZAC gene expression in gastric cancer cells in a time-and dose-dependant manner. The results of Hind Ⅲ/EcoRⅠ digestion and sequencing indicated that ZAC-shRNA expression vectors were constructed successfully. The ZAC mRNA level in the gastric cancer cells transfected with shRNA-ZAC/2 was decreased significantly (P<0.05), which were the gastric cancer cell lines in which ZAC gene was knock-down. The proliferation of the gastric cancer cell lines in which ZAC gene was knock-down was significantly higher than that of corresponding BGC823 and SGC7901 (P<0.05). After OCT incubation, the proliferation of BGC823 and SGC7901 were decreased obviously (P<0.05); however, that of the cells in which ZAC gene was knock-down did not exhibit significant changes (P>0.05). Conclusion OCT induces ZAC gene expression in gastric cancer cells in a time-and dose-dependant manner. ZAC gene may play an important role in the pathway of OCT inhibiting the proliferation of gastric cancer cells.

Key words: Octreotide, ZAC, Gastric cancer cell, Proliferation, RNA interference, Western blotting

DAI Wen DING Yi ZHANG Qin-xian*. Role of ZAC gene in the pathway of octreotide inhibiting proliferation of gastric cancer cells in vitro[J]. AAS, 2013, 44(4 ): 492-497.

References

［1］Abdollahi A. LOT1 (ZAC/PLAGL1) and its family members mechanisms and function［J］. J Cell Physiol, 2007, 210(1): 16-25.
［2］War SA, Kumar U. Coexpression of human somatostatin receptor-2 (SSTR2) and SSTR3 modulates antiproliferative signaling and apoptosis［J］. J Mol Signal, 2012, 7(1): 5-19.
［3] Zhang GZh, Ding Y, Liu J, et al. Correlation of ZAC with somatostatin and its receptor expression in gastric cancer tissues［J］. Acta Anatomica Sinaca, 2008, 39 (5): 703-707.(in Chinese)
张广政，丁一，刘键，等．胃癌组织中ZAC与生长抑素及生长抑素受体表达的相关性［J］．解剖学报，2008，39（5)：703-707. 
［4］Weckbecker G, Lewis I, Albert R, et al. Opportunities in somatostatin research: biological, chemical and therapeutic aspects［J］. Nat Rev Drug Discov, 2003, 2(12): 999-1017.［5］May P, May E. Twenty years of p53 research: structural and functional aspects of the p53 protein［J］. Oncogene, 1999,18 (2): 7621-7636.
［6］Shibui T, Higo Y, Tsutsui TW, et al. Changes in expression of imprinted genes following treatment of human cancer cell lines with nonmutagenic or mutagenic carcinogens［J］. Int J Oncol, 2008, 33(2): 351-360.
［7］Midorikawa Y, Yamamoto S, Ishikawa S, et al. Molecular karyotyping of human hepatocellular carcinoma using single-nucleotide polymorphism arrays［J］. Oncogene, 2006, 25(40): 5581-5590.
［8］Qi QG, Ma J, Da G, et al. Expression of zac in esophageal squamous cell carcinoma and its clinical significance［J］. Journal of the Fourth Military Medical University, 2007,28(19): 1793-1795.(in Chinese)
祁秋干,马军, 达嘎, 等.食管鳞癌中zac基因的表达及临床意义［J］. 第四军医大学学报, 2007,28(19)：1793-1795.
［9］Jarmalaite S, Laurinaviciene A, Tverkuviene J, et al. Tumor suppressor gene ZAC/PLAGL1: altered expression and loss of the nonimprinted allele in pheochromocytomas［J］. Cancer Genet, 2011, 204(7): 398-404.
［10］Valleley EM, Cordery SF, Carr IM, et al. Loss of expression of ZAC/PLAGL1 in diffuse large B-cell lymphoma is independent of promoter hypermethylation［J］. Genes Chromosomes Cancer, 2010, 49(5): 480-486.
［11］Theodoropoulou M, Tichomirowa MA, Sievers C, et al. Tumor ZAC1 expression is associated with the response to somatostatin analog therapy in patients with acromegaly［J］. Int J Cancer, 2009, 125(9): 2122-2126.
［12］Theodoropoulou M, Zhang J, Laupheimer S, et al. Octreotide, a somatastatin analogue, mediate its antiproliferative action in pituitary tumor cells by altering phosphatidylinotol 3-kinase signaling and inducing Zac1 expression［J］. Cancer Res, 2006, 66(3): 1576-1582.
［13］Wang CH, Tang CW, Liu CL, et al. Inhibitory effect of octreotide on gastric cancer growth via MAPK pathway［J］. World J Gastroenterol, 2003, 9(9): 1904-1908.
［14］El-Salhy M. Effects of octreotide, galanin and serotonin on a human gastric cancer cell line［J］. Oncol Rep, 2005, 13(5): 787-791.
［15］Theodoropoulou M, Stalla GK, Spengler D. ZAC1 target genes and pituitary tumorigenesis［J］. Mol Cell Endocrinol, 2010, 326(1-2): 60-65. 

[1]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[2]	WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen. Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia [J]. Acta Anatomica Sinica, 2024, 55(1): 3-9.
[3]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[4]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[5]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[6]	LÜ Hai-yan SHEN Xi-ya ZHAO Fu-sheng ZHU Mei . Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage [J]. Acta Anatomica Sinica, 2024, 55(1): 49-54.
[7]	ZHENG Li-ping LIU Xia DU Xiao-hua WU Ya-juan LIU Shan-shan . Expression and distribution of brain-derived neurotrophic factor in different cerebrum regions of yak and cattle [J]. Acta Anatomica Sinica, 2024, 55(1): 10-16.
[8]	HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe . Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method [J]. Acta Anatomica Sinica, 2024, 55(1): 120-124.
[9]	XU Ya-ping YU Yue-xin CHEN Jin-fu WANG Ke-ke GUO Zhikun . Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats [J]. Acta Anatomica Sinica, 2023, 54(6): 716-721.
[10]	ZHAO Wei-ming LI Xiao YU Guo-ying WANG Gai-ping CHANG Cui-fang . Effect of serine protease inhibitor Kazaltype 1 on the proliferation of hepatocellular carcinoma cell and its mechanism [J]. Acta Anatomica Sinica, 2023, 54(6): 695-702.
[11]	WANG Wen-juan DING Yu-tong SU Hao REN Jie SUN Yan-rong WANG Han-fei LU Jia-li ZHANG Lin-qian BAI Yu QIN Li-hua. Changes of Nogo-A expression in hippocampus and striatum of rats under low estrogen condition [J]. Acta Anatomica Sinica, 2023, 54(6): 620-627.
[12]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[13]	LIANG Pan-pan YU Ai-mei DU Jing KOU Wen-hui WANG Huan-huan SONG Ai-xia. Inhibitory effect of sodium ferulate on inflammatory response in migraine rats based on c-Jun N-terminal kinase/p38 mitogen-activated protein kinase signaling pathway [J]. Acta Anatomica Sinica, 2023, 54(6): 652-659.
[14]	XU Ying LIU Bin ZHENG Xing HE Zong-zhao. Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation [J]. Acta Anatomica Sinica, 2023, 54(6): 668-675.
[15]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.