Effects of small interfering RNA interference of connexin 37 on subcutaneous gastric tumours in mice

Jing,Yuanming; Guo,Suxia; Zhang,Xiaoping; Sun,Aijing; Tao,Feng; Ju,Haixing; Qian,Haixin

doi:10.3892/mmr.2014.2609

Effects of small interfering RNA interference of connexin 37 on subcutaneous gastric tumours in mice

Authors:
- Yuanming Jing
- Suxia Guo
- Xiaoping Zhang
- Aijing Sun
- Feng Tao
- Haixing Ju
- Haixin Qian
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Affiliations: Department of Gastrointestinal Surgery, Shaoxing People's Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, Zhejiang 312000, P.R. China, Department of Cardiology, The Affiliated People's Hospital of Nanjing Medical University in Wuxi and People's Hospital of Wuxi City, Wuxi, Jiangsu 214023, P.R. China, Department of Colorectal Surgery, Zhejiang Provincial Tumor Hospital, Hangzhou, Zhejiang 310022, P.R. China, Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: October 8, 2014 https://doi.org/10.3892/mmr.2014.2609
Pages: 2955-2960

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Abstract

The present study aimed to investigate the effects of small interfering (si)RNA interference of connexin 37 (Cx37) on subcutaneous gastric tumours in mice. Constructed lentiviruses carrying siRNA against Cx37 significantly knocked down Cx37 mRNA and protein expression in vitro. A total of 60 mice with gastric cancer were randomly divided into the Cx37 siRNA group, the mock‑siRNA group and the control group. Cx37 siRNA, mock‑siRNA and saline were separately injected (with the lentiviruses transfected into the gastric cancer cells). Following six weeks, the Cx37 mRNA expression, Cx37 protein expression and tumor apoptosis were detected using semiquantitative reverse transcription‑polymerase chain reaction, western blot analysis and terminal deoxynucleotidyl transferase‑mediated dUTP nick end labelling, respectively. Six weeks following lentiviral transfection, the Cx37 mRNA levels in the Cx37 siRNA group, mock‑siRNA group and saline group decreased to 42, 63 and 67%, respectively (P<0.05). The mock‑siRNA group demonstrated no significant change in Cx37 levels compared with the control group. Western blot analysis revealed lower Cx37 protein levels in the Cx37‑RNAi group than in the other groups (0.21±0.07 vs. 0.65±0.06 vs. 0.54±0.07), and that the apoptotic index of the Cx37‑RNAi group was higher than those of the mock‑siRNA and control groups (19.7±5.1 vs. 9.8±6.4 vs. 10.5±7.2%, 11.1±6.9; P<0.05). In conclusion, it was demonstrated that Cx37 siRNA is correlated with gastric cancer. Interference of Cx37 effectively reduces Cx37 mRNA and protein expression and promotes tumour apoptosis.

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1	Crew KD and Neugut AI: Epidemiology of gastric cancer. World J Gastroenterol. 12:354–362. 2006.
2	Li GX and Hu YF: The current status and perspective of laparoscopic gastrectomy for gastric cancer. Zhonghua Wai Ke Za Zhi. 51:389–392. 2013.(In Chinese).
3	Tsai JS, Lin MT, Wu MS, et al: PBMC expressed adiponectin mRNA is predictive of survival in patients with gastric cancer. Clin Chem Lab Med. 51:e313–316. 2013.PubMed/NCBI
4	Chong ML, Loh M, Thakkar B, Pang B, Iacopetta B and Soong R: Phosphatidylinositol-3-kinase pathway aberrations in gastric and colorectal cancer: meta-analysis, co-occurrence and ethnic variation. Int J Cancer. 134:1232–1238. 2014. View Article : Google Scholar : PubMed/NCBI
5	Li HJ, Che XM, Zhao W, et al: Diet-induced obesity promotes murine gastric cancer growth through a nampt/sirt1/c-myc positive feedback loop. Oncol Rep. 30:2153–2160. 2013.PubMed/NCBI
6	Völgyi B, Kovács-Oller T, Atlasz T, Wilhelm M and Gábriel R: Gap junctional coupling in the vertebrate retina: variations on one theme? Prog Retin Eye Res. 34:1–18. 2013.PubMed/NCBI
7	Marandykina A, Palacios-Prado N, Rimkutė L, Skeberdis VA and Bukauskas FF: Regulation of connexin36 gap junction channels by n-alkanols and arachidonic acid. J Physiol. 591:2087–2101. 2013. View Article : Google Scholar : PubMed/NCBI
8	Paul M, Wichter T, Gerss J, et al: Connexin expression patterns in arrhythmogenic right ventricular cardiomyopathy. Am J Cardiol. 111:1488–1495. 2013. View Article : Google Scholar : PubMed/NCBI
9	Foley TT and Ehrlich HP: Through gap junction communications, co-cultured mast cells and fibroblasts generate fibroblast activities allied with hypertrophic scarring. Plast Reconstr Surg. 131:1036–1044. 2013. View Article : Google Scholar
10	Tencé M, Ezan P, Amigou E and Giaume C: Increased interaction of connexin43 with zonula occludens-1 during inhibition of gap junctions by G protein-coupled receptor agonists. Cell Signal. 24:86–98. 2012.PubMed/NCBI
11	Krutovskikh V, Mironov N and Yamasaki H: Human connexin 37 is polymorphic but not mutated in tumours. Carcinogenesis. 17:1761–1763. 1996. View Article : Google Scholar : PubMed/NCBI
12	Good ME, Nelson TK, Simon AM and Burt JM: A functional channel is necessary for growth suppression by Cx37. J Cell Sci. 124:2448–2456. 2011. View Article : Google Scholar : PubMed/NCBI
13	Burt JM, Nelson TK, Simon AM and Fang JS: Connexin 37 profoundly slows cell cycle progression in rat insulinoma cells. Am J Physiol Cell Physiol. 295:C1103–C1112. 2008. View Article : Google Scholar : PubMed/NCBI
14	Simon KW, Roberts PC, Vespremi MJ, Manchen S and Schmelz EM: Regulation of beta-catenin and connexin-43 expression: targets for sphingolipids in colon cancer prevention. Mol Nutr Food Res. 53:332–340. 2009. View Article : Google Scholar : PubMed/NCBI
15	Cocciadiferro L, Miceli V, Kang KS, Polito LM, Trosko JE and Carruba G: Profiling cancer stem cells in androgen-responsive and refractory human prostate tumor cell lines. Ann NY Acad Sci. 1155:257–262. 2009. View Article : Google Scholar : PubMed/NCBI
16	Morel S, Burnier L, Roatti A, et al: Unexpected role for the human Cx37 C1019T polymorphism in tumour cell proliferation. Carcinogenesis. 31:1922–1931. 2010. View Article : Google Scholar : PubMed/NCBI
17	Jing YM, Guo SX, Zhang XP, Sun AJ, Tao F and Qian HX: Association between C1019T polymorphism in the connexin 37 gene and Helicobacter pylori infection in patients with gastric cancer. Asian Pac J Cancer Prev. 13:2363–2367. 2012. View Article : Google Scholar : PubMed/NCBI
18	Hou Q, Tang J, Wang Z, et al: Inhibitory effect of microRNA-34a on retinal pigment epithelial cell proliferation and migration. Invest Ophthalmol Vis Sci. 54:6481–6488. 2013. View Article : Google Scholar : PubMed/NCBI
19	Qin B, Chen Z, Jin W and Cheng K: Development of cholesteryl peptide micelles for siRNA delivery. J Control Release. 172:59–168. 2013.PubMed/NCBI
20	Li J and Mansmann UR: Modeling of non-steroidal anti-inflammatory drug effect within signaling pathways and miRNA-regulation pathways. PLoS One. 8:e724772013. View Article : Google Scholar : PubMed/NCBI
21	Yao Y, Charlesworth J, Nair V and Watson M: MicroRNA expression profiles in avian haemopoietic cells. Front Genet. 4:1532013.PubMed/NCBI
22	Reinhard K, Rougier JS, Ogrodnik J and Abriel H: Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Na v1.5 expressed in HEK293 cells. F1000Res. 2:482013.PubMed/NCBI
23	Chen HJ, Zhu Z, Wang XL, et al: Expression of huCdc7 in colorectal cancer. World J Gastroenterol. 19:3130–3133. 2013. View Article : Google Scholar : PubMed/NCBI
24	Zhu Q, Gu L, Wang Y, et al: The role of alpha-1 and alpha-2 adrenoceptors in restraint stress-induced liver injury in mice. PLoS One. 9:e921252014. View Article : Google Scholar : PubMed/NCBI
25	Kuphal S, Wallner S and Bosserhoff AK: Impact of LIF (leukemia inhibitory factor) expression in malignant melanoma. Exp Mol Pathol. 95:156–165. 2013. View Article : Google Scholar : PubMed/NCBI
26	Billingsley CN, Allen JR, Baumann DD, et al: Non-trisomic homeobox gene expression during craniofacial development in the Ts65Dn mouse model of Down syndrome. Am J Med Genet A. 161:1866–1874. 2013. View Article : Google Scholar : PubMed/NCBI
27	Ohnishi S, Ma N, Thanan R, et al: DNA damage in inflammation-related carcinogenesis and cancer stem cells. Oxid Med Cell Longev. 2013:3870142013. View Article : Google Scholar : PubMed/NCBI
28	Kwak HW, Choi IJ, Cho SJ, et al: Characteristics of gastric cancer according to Helicobacter pylori infection status. J Gastroenterol Hepatol. 29:1671–1677. 2014.
29	Hosogi S, Kusuzaki K, Inui T, Wang X and Marunaka Y: Cytosolic chloride ion is a key factor in lysosomal acidification and function of autophagy in human gastric cancer cell. J Cell Mol Med. 18:1124–1133. 2014. View Article : Google Scholar : PubMed/NCBI
30	Oo HZ, Sentani K, Sakamoto N, et al: Overexpression of ZDHHC14 promotes migration and invasion of scirrhous type gastric cancer. Oncol Rep. 32:403–410. 2014.PubMed/NCBI