Regulation of FN1 degradation by the p62/SQSTM1-dependent autophagy-lysosome pathway in HNSCC / 国际口腔科学杂志·英文版

Epithelial-mesenchymal transition (EMT) is involved in both physiological and pathological processes. EMT plays an essential role in the invasion, migration and metastasis of tumours. Autophagy has been shown to regulate EMT in a variety of cancers but not in head and neck squamous cell carcinoma (HNSCC). Herein, we investigated whether autophagy also regulates EMT in HNSCC. Analyses of clinical data from three public databases revealed that higher expression of fibronectin-1 (FN1) correlated with poorer prognosis and higher tumour pathological grade in HNSCC. Data from SCC-25 cells demonstrated that rapamycin and Earle's balanced salt solution (EBSS) promoted autophagy, leading to increased FN1 degradation, while 3-methyladenine (3-MA), bafilomycin A1 (Baf A1) and chloroquine (CQ) inhibited autophagy, leading to decreased FN1 degradation. On the other hand, autophagic flux was blocked in BECN1 mutant HNSCC Cal-27 cells, and rapamycin did not promote autophagy in Cal-27 cells; also in addition, FN1 degradation was inhibited. Further, we identified FN1 degradation through the lysosome-dependent degradation pathway using the proteasome inhibitor MG132. Data from immunoprecipitation assays also showed that p62/SQSTM1 participated as an autophagy adapter in the autophagy-lysosome pathway of FN1 degradation. Finally, data from immunoprecipitation assays demonstrated that the interaction between p62 and FN1 was abolished in p62 mutant MCF-7 and A2780 cell lines. These results indicate that autophagy significantly promotes the degradation of FN1. Collectively, our findings clearly suggest that FN1, as a marker of EMT, has adverse effects on HNSCC and elucidate the autophagy-lysosome degradation mechanism of FN1.

Osteogenic induction and identification of mammary stem cells derived from a Holstein dairy cow / 中国组织工程研究

BACKGROUND: The isolated and purified mammary stem cells from Holstein dairy cows have been induced to differentiate into nerve cells, adipocytes and osteoblasts, further enriching their multi-directional differentiation potential, which is of great significance for the study on mammary stem cells. OBJECTIVE: To investigate the osteogenic potential of mammary stem cells from Holstein dairy cows. METHODS: Mammary stem cells from Holstein dairy cows were isolated and purified, which were provided by the Key Laboratory of Biological Manufacturing of Inner Mongolia Autonomous Region. The fourth generation of mammary stem cells were cultured for 21 days in an osteogenic induction medium, and identified by alizarin red staining and RT-PCR. RESULTS AND CONCLUSION: After osteogenic induction, the cell edge was indistinct, and there were some granular calcified nodules in the cells. The induced cells were positive for alizarin red staining. The positive expressions of Alpl and Bglap in the induced osteoblasts were detected, indicating that the mammary stem cells from Holstein dairy cows have the potential to differentiate into osteoblasts.

Osteogenic induction and identification of mammary stem cells derived from a Holstein dairy cow / 中国组织工程研究

BACKGROUND: The isolated and purified mammary stem cells from Holstein dairy cows have been induced to differentiate into nerve cells, adipocytes and osteoblasts, further enriching their multi-directional differentiation potential, which is of great significance for the study on mammary stem cells. OBJECTIVE: To investigate the osteogenic potential of mammary stem cells from Holstein dairy cows. METHODS: Mammary stem cells from Holstein dairy cows were isolated and purified, which were provided by the Key Laboratory of Biological Manufacturing of Inner Mongolia Autonomous Region. The fourth generation of mammary stem cells were cultured for 21 days in an osteogenic induction medium, and identified by alizarin red staining and RT-PCR. RESULTS AND CONCLUSION: After osteogenic induction, the cell edge was indistinct, and there were some granular calcified nodules in the cells. The induced cells were positive for alizarin red staining. The positive expressions of Alpl and Bglap in the induced osteoblasts were detected, indicating that the mammary stem cells from Holstein dairy cows have the potential to differentiate into osteoblasts.