Oma.19 Furthermore, LGR5 has been recognized as a CSC marker
Oma.19 Additionally, LGR5 has been recognized as a CSC marker for colorectal cancers.20 Our preceding study showed that LGR5 was progressively expressed in cervical carcinogenesis and promoted the proliferation of cervical cancer cells at the same time as tumor formation by potentiating the Wnt/-catenin pathway.11 Therefore, we hypothesized that LGR5 could possibly contribute to cervical carcinogenesis, recurrence and metastasis by keeping the stemness of cervical CSCs. In this study, we used typical functional IFN-beta Protein medchemexpress assays and fluorescence activated cell sorting (FACS) to analyze the properties of cervical CSCs with different levels of LGR5 expression. The outcomes indicated that a subpopulation of human cervical cancer cells with elevated LGR5 expression possesses enhanced self-renewal capacity, differentiation possible and tumorigenicity. Furthermore, we found that the elevated tumorsphere-forming efficiency induced by LGR5 could be regulated via the inhibition or activation of the Wnt/-catenin pathway in cervical cancer cells. Our dataDepartment of Reproductive Medicine, the very first Affiliated Hospital, Xi’an Jiaotong University Health-related School, Xi’an 710061, China; 2Department of Pharmacology, Shannxi Provincial Tumor Hospital, Xi’an 710061, China; 3Department of Obstetrics and Gynecology, the Second Affiliated Hospital, Xi’an Jiaotong University Health-related School, Xi’an 710061, China and 4Division of Cancer Stem Cell Investigation, Important Laboratory of Atmosphere and Genes Related to Diseases, Ministry of Education, Xi’an Jiaotong University Medical School, Xi’an 710061, China Corresponding author: P-S Zheng, Department of Reproductive Medicine, The very first Affiliated Hospital of your Medical College, Xi’an Jiaotong University, 76 West Yanta Road, Xi’an 710061, China. Tel: +86 29 82657874; Fax: +86 29 85324013; E-mail: [email protected] 5 These authors contributed equally to this workReceived 03.5.17; revised 07.7.17; accepted ten.7.17; Edited by A StephanouLGR5 promotes CSC traits in cervical cancer H-Z Cao et alFigure 1 Overexpression and knockdown of LGR5 in human cervical cancer cell lines. (a) Immunohistochemical staining displaying LGR5 expression in LGR5-overexpressing and LGR5-knockdown SiHa and HeLa cells, scale bar, ten m. (b) A western blot assay was employed to characterize the expression of LGR5 in LGR5-overexpressing and LGR5-knockdown SiHa and HeLa cells. (c) The expression of LGR5 in HeLa, SiHa cells was measured by western blot. (d) LGR5 expresssion was analyzed by flow cytometry. AcGFP: green fluorescent protein for manage; LGR5: overexpression for LGR5; shLGR5: shRNA for LGR5; shControl: shRNA for control. Values are shown as the imply sirtuininhibitorS.D. Po0.05; Po0.indicated that LGR5 includes a essential PDGF-BB Protein web oncogenic role through promoting CSC traits in cervical cancer. Results Elevated LGR5 enhances the self-renewal capacity of cervical cancer cells. As a CSC marker, LGR5 has been shown to be progressively expressed in cervical carcinogenesis and to market cancer cell proliferation and tumor formation. To additional investigate the mechanism involved in tumor promotion by LGR5 in cervical cancer, stable LGR5overexpressing cells (SiHa-LGR5 and HeLa-LGR5) and stable LGR5-knockdown cells (SiHa-shLGR5 and HeLashLGR5) have been established in cervical cancer cell lines. The expression of LGR5 in these cell lines was examined by immunohistochemistry (IHC), western blot and flow cytometry. As shown in Figure 1a, higher expression of LGR5 was detected in SiHa-LGR5 and HeLa-.