ABSTRACT
OBJECTIVE@#To explore the regulatory mechanism of human hepatocyte apoptosis induced by lysosomal membrane protein Sidt2 knockout.@*METHODS@#The Sidt2 knockout (Sidt2-/-) cell model was constructed in human hepatocyte HL7702 cells using Crispr-Cas9 technology.The protein levels of Sidt2 and key autophagy proteins LC3-II/I and P62 in the cell model were detected using Western blotting, and the formation of autophagosomes was observed with MDC staining.EdU incorporation assay and flow cytometry were performed to observe the effect of Sidt2 knockout on cell proliferation and apoptosis.The effect of chloroquine at the saturating concentration on autophagic flux, proliferation and apoptosis of Sidt2 knockout cells were observed.@*RESULTS@#Sidt2-/- HL7702 cells were successfully constructed.Sidt2 knockout significantly inhibited the proliferation and increased apoptosis of the cells, causing also increased protein expressions of LC3-II/I and P62(P < 0.05) and increased number of autophagosomes.Autophagy of the cells reached a saturated state following treatment with 50 μmol/L chloroquine, and at this concentration, chloroquine significantly increased the expressions of LC3B and P62 in Sidt2-/- HL7702 cells.@*CONCLUSION@#Sidt2 gene knockout causes dysregulation of the autophagy pathway and induces apoptosis of HL7702 cells, and the latter effect is not mediated by inhibiting the autophagy-lysosomal pathway.
Subject(s)
Humans , Lysosomal Membrane Proteins/metabolism , Autophagy , Apoptosis , Hepatocytes , Lysosomes/metabolism , Chloroquine/pharmacology , Nucleotide Transport Proteins/metabolismABSTRACT
Objective To explore the clinical application value of prognostic nutritional index (PNI)for predicting overall survival(OS)in patients with advanced colorectal cancer.Methods Seventy-two patients with histologically confirmed colorectal cancer were enrolled in this study, and their clinical and laboratory data were reviewed.The PNI was calculated, and univariate and multivariate analysis was used to identify the potential prognostic factors for advanced colorectal cancer. Results PNI of the 72 colorectal cancer patients was 45.07 ± 5.98.PNI was significantly associated with age, weight loss and pleural effusion (P < 0.05). However, PNI showed no correlation with sex, clinical stage, smoking, bloody stool, abdominal mass, intestinal obstruction, histological type, radiotherapy and KPS scores (P>0.05).The median OS of the 72 patients was 12.9 months.The medium OS in the higher PNI group (PNI≥45.07)and lower PNI group(PNI≤45.07)was 15.7 months and 11.2 months, respectively.The 1-year survival rates were 78.4% and 60.5%, and 2-year survival rates were 53.1% and 20.8%, respectively(P<0.01).Univariate analysis showed that PNI, ages, and weight loss were related to the OS of the advanced colorectal cancer(P<0.05).Multivariate analysis identified PNI was an independent prognostic factor for OS of advanced colorectal cancer (P < 0.01). Conclusions PNI can be easily calculated, and may be used as a relatively new prognostic indicator for advanced colorectal cancer in clinical practice.