Jun Yin

Spartin Promotes Smurf1‐Mediated Ubiquitination Modification of YWHAZ to Inhibit Cisplatin Resistance in Ovarian Cancer

ABSTRACT Cisplatin (DDP) remains the commonly used chemotherapeutic drug for ovarian cancer (OV); however, DDP resistance poses a great challenge to the outcomes of patients. This work investigated the biological function and mechanism of Spartin in DDP resistance of OV. The growth and apoptosis of DDP‐resistant OV cells were assessed by CCK‐8, colony formation, and flow cytometry, respectively. Autolysosome fusion was observed by immunofluorescent staining of LC3 and LAMP2. The interaction between E3 ligase Smurf1 and YWHAZ or Spartin protein, and the ubiquitination level of YWHAZ were determined by Co‐IP assay. Expression levels of autophagy or apoptosis‐related markers were measured by RT‐qPCR, western blotting, and immunohistochemistry. DDP resistance was assessed by xenograft tumor experiments in vivo. We found that Spartin expression was lower, while YWHAZ expression was higher in DDP‐resistant OV samples and cells. Lower expression of Spartin indicated a poorer survival rate of OV patients. In addition, overexpression of Spartin sensitized OV cells to DDP and repressed autophagy. Moreover, Spartin bound to Smurf1 to promote Smurf1‐mediated ubiquitination and degradation of YWHAZ, restraining autophagy and DDP resistance. Overexpression of YWHAZ counteracted the effects of Spartin against DDP resistance by promoting autophagy. In conclusion, Spartin‐induced Smurf1‐mediated ubiquitination modification of YWHAZ to inactivate autophagy, thereby increasing the sensitivity of OV cells to DDP. Our findings suggest that Spartin‐combined therapy might act as an effective approach to fight against DDP resistance in OV.

circ_C20orf11 enhances DDP resistance by inhibiting miR-527/YWHAZ through the promotion of extracellular vesicle-mediated macrophage M2 polarization in ovarian cancer

Ovarian cancer is a fatal gynecologic tumor, and conventional treatment is mainly limited by chemoresistance. The mechanism contributing to chemoresistance in ovarian cancer has yet to be established. This study aimed to investigate the specific role of circ_C20orf11 in regulating chemoresistance to cisplatin (DDP)in ovarian cancer. We first established two DDP-resistant ovarian cancer cell lines. Then, we identified the effect of circ_C20orf11 on specific cellular characteristics (proliferation, apoptosis, DDP resistance) via a series of experiments. The binding sites between circ_C20orf11 and miR-527 and between miR-527 and YWHAZ were predicted using a bioinformatics tool and confirmed with a dual-luciferase reporter assay. Furthermore, extracellular vesicles (EVs) derived from DDP-resistant cell lines were identified, and the effect of EVs on macrophage polarization was examined. circ_C20orf11 was upregulated in ovarian cancer. Increased circ_C20orf11 expression enhanced DDP resistance and cell proliferation and reduced cell apoptosis in DDP-resistant cell lines after DDP treatment by sponging miR-527 and promoting YWHAZ expression. In addition, we found that DDP-resistant cell-derived EVs can induce macrophage M2 polarization, whereas silencing of circ_C20orf11 inhibited EV-induced macrophage M2 polarization. Consistent with these results, silencing of circ_C20orf11 enhanced sensitivity to DDP in vivo. Importantly, we proved that circ_C20orf11 expression was upregulated in EVs extracted from the serum of DDP-resistant patients. Our study demonstrated that silencing circ_C20orf11 sensitizes ovarian cancer to DDP by promoting miR-527/YWHAZ signaling and EV-mediated macrophage M2 polarization.