Lan Wang's research while affiliated with First Affiliated Hospital of China Medical University and other places

Aim To determine the role of claudin-3 in cancer stemness in nonsquamous non-small-cell lung carcinoma (NSCLC). Materials & methods In vitro/vivo extreme limiting dilution analysis and the side population assay were used to investigate the role of claudin-3 in regulating cancer stemness in nonsquamous NSCLC. Results & conclusion Claudin-3 depletion decreased the formation rates of spheres and tumors and increased cisplatin sensitivity. Claudin-3 was also identified as one downstream target of estrogen receptor-α in regulating cancer stemness. Moreover, targeting CLDN-3 transcription by small molecules including withaferin A, estradiol and fulvestrant suppressed cancer stemness and reversed chemoresistance. These results demonstrated claudin-3 is one positive regulator of cancer stemness in nonsuqamous NSCLC.

Effect of double knockdown of CXADR and LNX1 on the percentage of SP. SP analysis was performed using BD Aria software and was analyzed using FlowJo 7.6.1 software. (TIF)

Cancer stem-like cells (CSCs) have been reported to play major roles in tumorigenesis, tumor relapse, and metastasis after therapy against colorectal carcinoma (CRC). Therefore, identification of colorectal CSC regulators could provide promising targets for CRC. Ligand-of-Numb protein X1 (LNX1) is one E3 ubiquitin ligase which mediates the ubiquitination and degradation of Numb. Although several studies indicate LNX1 could be a potential suppressor of cancer diseases, the functions of LNX1 in mediating cancer stemness remain poorly understood. In this study, LNX1 was identified as a negative regulator of cancer stemness in CRC, which was downregulated in colonospheres or side population (SP) cells. Furthermore, the coxsackievirus and adenovirus receptor (CXADR) was found to be one critical downstream mediator of cancer stemness regulated by LNX1. Interestingly, the anti-breast cancer drug tamoxifen was found to be an agonist of LNX1 and suppress cancer stemness in CRC. In sum, this study provided the evidences that LNX1 signaling plays important roles in regulating the stemness of colon cancer cells.

Analysis of the function of LNX1 in mediating cancer stemness in colorectal carcinoma cell line Colo205. (A) Effect of LNX1 knockdown on the percentage of SP in Colo205. (B) Effect of LNX1 knockdown on the capacities of colonosphere formation (n = 8 per group). (C) Effect of LNX1 knockdown on the rates of colonosphere formation (p value was calculated using the online ELDA software). (D) The efficiency of LNX1 knockdown using two shLNX1 constructs. Data from triplicates are presented as the mean±SD, *P<0.05, **P<0.01, ***P<0.001. (TIF)

Histological analysis of HT29 xenograft tumors. Tumor sections were stained with hematoxylin and eosin (H & E). (TIF)

Screening of downstream targets of LNX1. SP analysis was performed using BD Aria software and was analyzed using FlowJo 7.6.1 software. (TIF)