Wiskot-Aldrich syndrome-like lower expressed in colon cancer tissues and inhibited the stemness of human colon cancer / 解剖学报

ABSTRACT

Objective To explore the expression of Wiskot￣Aldrich syndrome-like(WASL)in colon cancer tissues and its function on colon cancer stemness. Methods The UALCAN database was used to analyze the expression of WASL in colon cancer tissues and its correlation among the gender, lymphnode stage and metastasis in colon cancer patients. Immunohistofluorescence and immunocyte fluorescence were used to verify the date from UALCAN. Western blotting was used to detect the expression and transfection efficiency of WASL in human normal colorectal mucosal cells (FHC cells) and colon cancer cells HCT-116, SW-480. Sphere formation assay and colony formation were used to detect the effect of WASL on the stemness of colon cancer. Hypoxia assay, serum deprivation, drug resistance assay were used to detect the stemness of colon cancer cells. Results WASL was lower expressed in colon cancer tissues (P<0. 05). WASL was lower expressed in colon cancer tissues than peri-cancer tissues. WASL was obviously lower expressed in HCT-116 and SW-480 than FHC cells. WASL overexpression stable cells were constructed suscessfully in HCT-116 and SW-480.Real-time PCR showed WASL obviously down-regulated the expression of cancer stemcells (CSCs) related markers [octamer binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2), Lin28, krüppel-like factor (KLF)]. Sphere formation assay and colony formation assay showed WASL could inhibited the sphere formation and colony formation. Hypoxia assay and serum deprivation showed WASL inhibited the resistance of cancer cells to hypoxia and serum deprivation. Drug resistance assay showed WASL inhibit the resistance of cancer cells to 5-fluorouracil(5-FU). Conclusion WASL is lower expressed in colon cancer tissues and cells, and is related with poor prognosis of colon cancer. Overexpression of WASL can inhibit the stemness of colon cancer cells.