Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there is still a lack of high-throughput methods for screening protein regulators of LLPS in target proteins. Here, we developed a CRISPR/Cas9-based screening method to identify protein phase separation regulators by integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly developed method, we screened the RNA-binding proteins that regulate PABPN1 phase separation and identified the tumor suppressor QKI as a promoter of PABPN1 phase separation. Furthermore, QKI exhibits decreased expression levels and diminished nuclear localization in colorectal cancer cells, resulting in reduced PABPN1 phase separation, which, in turn, promotes alternative polyadenylation (APA), cell proliferation, and migration in colorectal cancer.
Humans ; Colorectal Neoplasms/genetics* ; RNA-Binding Proteins/genetics* ; Poly(A)-Binding Protein I/genetics* ; CRISPR-Cas Systems ; Flow Cytometry ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

To investigate the correlation between soluble resistance-related calcium-binding protein (Sorcin) and chemoresistance or overall survival in patients with ovarian cancer.
 Methods: We detected the expression of Sorcin in 27 cases of chemoresistant ovarian cancer tissue and 37 cases of sensitive ovarian cancer tissue by immunohistochemistry, and analyzed the relationship between the protein and clinicopathological features or chemoresistance of ovarian cancer. Log-rank test was used to analyze the single factor impact on overall survival, Kaplan-Meier analysis was used to describe survival curve, and Cox proportional hazard model was used for multivariate analysis.
 Results: The immunoreactive scores for Sorcin in chemoresistant ovarian cancer tissues were higher than those in the sensitive ovarian cancer tissues (P<0.001). The levels of Sorcin inovarian cancer tissue did not show statistical significance with different ages, tumor stages, classifications, tissue types, degrees of ascites, omentums, and tumor metastases (P>0.05). The correlation between Sorcin and overall survival in resistant and sensitive ovarian cancer groups was not statistically significant (P>0.05), while there was a negative correlation between the expression of Sorcin and the overall survival of total cases (r=-0.326, P<0.05). Log-rank test showed that the drug resistance factor had a distinct impact on overall survival (P<0.001), and the Sorcin expression had an impact on overall survival (P<0.05). However, correlation between overall survival and the ages, ascites, omentum carcinoma, pathological types, pathological grade or FIGO staging was not significant (P>0.05). Cox proportional hazard model showed that drug resistance had a significant effect on overall survival (P<0.001), with a relative risk at 8.635, and the survival curve of the ovarian cancer sensitive group was obviously superior to that of ovarian cancer drug resistance group.
 Conclusion: Sorcin may be associated with drug resistance in ovarian cancer. The expression of Sorcin is correlated with the overall survival. The lower the Sorcin expression, the longer the survival time. Chemoresistance may act as an important independent prognostic factor for the poor prognosis for ovarian cancer.
Biomarkers, Tumor ; Carcinoma, Ovarian Epithelial ; Drug Resistance, Neoplasm ; Female ; Humans ; Kaplan-Meier Estimate ; Neoplasm Staging ; Ovarian Neoplasms ; Prognosis

Objective To investigate the hearing outcome efficiency of tympanostomy tubes and when combined with adenoidectomy for otitis media with effusion treatment. Methods A systematic literature review of tympanostomy tubes and tympanostomy tubes combined with adenoidectomy were performed using PubMed, EMBASE,the Cochrane Library,MEDLINE,all of the included studies were randomized controlled trial. The quality of included studies was evaluated according to the Risk of Bias Table of the Cochrane Handbook. Data were analyzed with Review Manager 5.02 software. Heterogeneity was examined,and forest plot was drawn. Results All of the included 4 studies were randomized controlled trials. At 6 month follow-up patients,the tym-panostomy tubes and tympanostomy tubes combined with adenoidectomy had no difference in hearing improvement (WMD = 1.06,95%CI-0.18 ~ 2.29,P = 0.09). At 12 month follow-up patients,the tympanostomy tubes com-bined with adenoidectomy were better than tympanostomy tubes alone(WMD = 3.03,95%CI 0.91 ~ 5.14,P =0.005). Conclusions The meta analysis result reveals that tympanostomy tubes combined with adenoidectomy have an advantage in long term follow up time for hearing improvement in children's otitis media with effusion treatment.