PMID- 31182056 OWN - NLM STAT- MEDLINE DCOM- 20191223 LR - 20200225 IS - 1471-2407 (Electronic) IS - 1471-2407 (Linking) VI - 19 IP - 1 DP - 2019 Jun 10 TI - Deletion of sorting nexin 27 suppresses proliferation in highly aggressive breast cancer MDA-MB-231 cells in vitro and in vivo. PG - 555 LID - 10.1186/s12885-019-5769-z [doi] LID - 555 AB - BACKGROUND: Sorting Nexin 27 (SNX27) belongs to a family of sortin nexins and possesses a unique binding domain at the C-terminus which mediates protein-protein interaction in intracellular trafficking, membrane remodeling, organelle motility, and tight junctions. However, its role in cancer development, especially in vivo, remains largely unknown. METHODS: We have generated a stable SNX27 knockdown clone in a highly aggressive breast cancer cell line MDA-MB-231 using an inducible lentiviral shRNA system. Cell migration and proliferation of SNX27 knockdown (KD) cells were compared with wild-type (WT) cells by MTT and wound healing assay, respectively. The differences in colony formation between SNX27-KD and WT cells were detected by soft agar culture and matrigel 3D culture. Furthermore, tumor growth was examined in a xenograft nude mouse model using SNX27-KD and WT MDA-MB-231 cells. The critical EMT (epithelial-mesenchymal transition) regulators were examined in vitro and in vivo. RESULTS: The wound healing assay showed that SNX27 knockdown significantly decreased cell motility and proliferation. Colony formation in soft agar showed that the SNX27 knockdown cells formed significantly fewer and smaller colonies than the parental MDA-MB-231 cells. Western blots and immunostaining showed that knockdown of SNX27 led to increased expression of E-cadherin and beta-catenin proteins, which facilitate adhesion formation and reverse EMT. EMT is a cellular program that allows polarized, immotile epithelial cells to convert to motile mesenchymal cells, promoting carcinoma invasion. The expression levels of Vimentin, the transcription factor of EMT, and tight junction protein Claudin-5, were significantly diminished in the SNX27 knockdown cells. The expression of PCNA, the cell proliferation marker, was increased in SNX27-KD cells transfected with E-cadherin siRNA. In a xenograft nude mouse model, we found that knockdown of SNX27 significantly inhibited tumor growth. The tumors from mice with SNX27-KD cells showed less proliferation compared to tumors from mice injected with wildtype cells. The increase in E-cadherin and beta-catenin and decrease in Vimentin and Claudin-5 were observed in tumors of mice injected with SNX27-KD cells. CONCLUSIONS: Our data have demonstrated that SNX27 plays a crucial role in tumor growth in vitro and in vivo. FAU - Zhang, Jilei AU - Zhang J AD - Division of Gastroenterology and Hepatology, Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, MC716, Chicago, IL, 60612, USA. FAU - Li, Kendy AU - Li K AD - Liberal Arts & Human Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA. FAU - Zhang, Yongguo AU - Zhang Y AD - Division of Gastroenterology and Hepatology, Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, MC716, Chicago, IL, 60612, USA. FAU - Lu, Rong AU - Lu R AD - Division of Gastroenterology and Hepatology, Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, MC716, Chicago, IL, 60612, USA. FAU - Wu, Shaoping AU - Wu S AD - Department of Biochemistry, Rush University, Chicago, IL, USA. FAU - Tang, Jingrong AU - Tang J AD - Section of Inflammation and Cardiometabolic Diseases, National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health, Bethesda, MD, USA. FAU - Xia, Yinglin AU - Xia Y AD - Division of Gastroenterology and Hepatology, Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, MC716, Chicago, IL, 60612, USA. FAU - Sun, Jun AU - Sun J AUID- ORCID: 0000-0001-7465-3133 AD - Division of Gastroenterology and Hepatology, Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, MC716, Chicago, IL, 60612, USA. junsun7@uic.edu. LA - eng GR - R01DK105118-01/Foundation for the National Institutes of Health/ GR - R01DK114126/Foundation for the National Institutes of Health/ GR - DOD CDMRP log No BC160450P1/U.S. Department of Defense/ PT - Journal Article DEP - 20190610 PL - England TA - BMC Cancer JT - BMC cancer JID - 100967800 RN - 0 (Cadherins) RN - 0 (RNA, Small Interfering) RN - 0 (SNX27 protein, human) RN - 0 (Sorting Nexins) SB - IM MH - Animals MH - Breast Neoplasms/*genetics MH - Cadherins/genetics/metabolism MH - Cell Movement MH - Cell Proliferation MH - Epithelial-Mesenchymal Transition MH - Female MH - Gene Expression Regulation, Neoplastic MH - Humans MH - MCF-7 Cells MH - Mice MH - Mice, Nude MH - RNA, Small Interfering/genetics MH - Sorting Nexins/*genetics MH - Xenograft Model Antitumor Assays PMC - PMC6558813 OTO - NOTNLM OT - Breast cancer OT - Cell adhesion OT - Cell junctions OT - Epithelial-mesenchymal transition OT - Proliferation OT - Sorting nexin 27 COIS- The authors declare that they have no competing interests. EDAT- 2019/06/12 06:00 MHDA- 2019/12/24 06:00 PMCR- 2019/06/10 CRDT- 2019/06/12 06:00 PHST- 2018/11/02 00:00 [received] PHST- 2019/05/29 00:00 [accepted] PHST- 2019/06/12 06:00 [entrez] PHST- 2019/06/12 06:00 [pubmed] PHST- 2019/12/24 06:00 [medline] PHST- 2019/06/10 00:00 [pmc-release] AID - 10.1186/s12885-019-5769-z [pii] AID - 5769 [pii] AID - 10.1186/s12885-019-5769-z [doi] PST - epublish SO - BMC Cancer. 2019 Jun 10;19(1):555. doi: 10.1186/s12885-019-5769-z.