The morbidity of oral cancer is high in the world. Oridonin is a traditional Chinese medicine that can effectively inhibit oral squamous cell carcinoma (OSCC) growth, but its mechanism remains unclear. Our previous data showed that oridonin inhibited CAL-27 cell proliferation and promoted apoptosis. Herein, we explored the mechanism and target of oridonin in human OSCC through RNA sequencing and integration of multiple bioinformatics analysis strategies. Differences in gene expression can be analyzed with RNA sequencing. Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), gene set enrichment analysis (GSEA), Disease Ontology (DO), and other enrichment analyses were used to evaluate differentially expressed genes (DEGs). Protein–protein interaction (PPI) networks were built via the STRING database. It was found that tumor necrosis factor (TNF) signaling pathway, cytokine–cytokine receptor interaction, and nuclear factor-kappa B (NF-kappaB) signaling pathway were associated with the therapeutic effects of oridonin in OSCC. Three key genes (BIRC3, TNFSF10, and BCL6) were found to associate with cell apoptosis in OSCC cells treated with oridonin. Quantitative PCR assays verified the expression of apoptosis-related DEGs: TNFSF10, BIRC3, AIFM2, BCL6, BCL2L2, and Bax. Western blots were employed for verifying proteins expression associated with DEGs: cleaved caspase 3, Bax, Bcl-w, anti-cIAP2, and anti-TRAIL. In conclusion, our findings reveal the molecular pathways and targets by which oridonin can treat and induce cytotoxic effects in OSCC: by affecting the signaling including TNF, NF-κB, and cytokine-cytokine receptor interaction and by regulating the key gene BIRC3, TNFSF10, and BCL6. It should be noted that further clinical trial validation is very necessary. Combined with current research trends, our existing research may provide innovative research drugs for the treatment of OSCC.

Background: Presenilin enhancer-2 (PSENEN, PEN-2), one of the four components of the γ-secretase complex, has been increasingly revealed to be significant in cancer types such as pancreatic cancer, gastric cancer, and breast cancer. However, the pan-cancer clinical relevance of PSENEN remains unclear. Methods: Raw data on PSENEN expression in normal and cancer tissues were obtained from The Cancer Genome Atlas (TGCA) database and Genotype-Tissue Expression Project (GTEx). The difference of PSENEN expression was analyzed using data from the TCGA repository and TIMER2 database. Meanwhile, Cox regression analysis and KM plotter were used to analyze the pan-cancer prognostic significance of PSENEN. We also analyzed the correlation between PSENEN expression and tumor immune infiltration using the TIMER and XCELL algorithms. Moreover, we used the TISIDB database to determine PSENEN expression in different immune and molecular subtypes of human cancers. Pan-cancer analysis of genetic alteration of PSENEN was performed using online tools such as cBioPortal and UALCAN. Finally, six Gene Expression Omnibus datasets from the Gene Expression Profiling Interactive Analysis database were used to validate the expression level of PSENEN in lung adenocarcinoma (LUAD). Results: Contrary to nonmalignant tissues, the expression level of PSENEN was significantly upregulated in the cancerous tissues in 22 cancer types. Elevated PSENEN expression was correlated with worse overall survival in 7 cancer types and with worse disease-specific survival in 8 of them. By using xCell algorithm, TIMER algorithm, and Spearman's correlation analysis, we found that PSENEN expression was closely correlated with the infiltration of immune cells across all cancer types. Moreover, aberrant PSENEN expression was associated with 60 immune checkpoint pathway-related genes, microsatellite instability, and tumor mutation burden in various cancer types. Besides, PSENEN expression was significantly associated with different immune subtypes and molecular subtypes of various human cancers. Notably, ovarian epithelial tumor samples demonstrated the highest frequency of PSENEN mutation among all cancer types. Conclusions: Our pan-cancer analysis demonstrated that PSENEN might serve as a prognostic biomarker and revealed the importance of an in-depth understanding of the oncogenic role of PSENEN in various malignancies.

e21058 Background: Intratumoral heterogeneity (ITH) is associated with the poor prognosis in a variety of solid tumors. However, the use of ITH as a predictive biomarker for immunotherapy in patients with advanced non-small cell lung cancer (NSCLC) is still lacking. Methods: We used the Bioconductor R package Maftools to analyze somatic variants of NSCLC from an independent cohort (Hellmman CM012 study cohort) with data from 75 patients with advanced NSCLC. We then used a mutant-allele tumor heterogeneity (MATH) algorithm to measure ITH and analyzed the correlation with immunogenic marker, and the prognostic effect of ITH on immunotherapy. Results: The objective response rate (ORR) and durable clinical benefit (DCB) of immunotherapy was 36.5% and 53.8% for the patients with low ITH (ITH-L), and 21.7% and 39.1% for the high ITH (ITH-H) subgroup (P = 0.0427, P = 0.3180). The ITH-L patients had significantly improved median progression free survival (mPFS) than the ITH-H group (7.98 versus 2.60 months, hazard ratio (HR) = 1.88, 95%CI 1.06−3.34, P = 0.0280). The ORR and DCB of immunotherapy was 45.9% and 59.5% for the patients with high TMB (TMB-H), and 18.4% and 39.5% for the low TMB (TMB-L) subgroup (P = 0.0354, P = 0.1077). The TMB-H patients had significantly better mPFS than the TMB-L group (8.12 versus 3.78 months, HR = 0.49, 95%CI 0.28−0.87, P = 0.0130). A pooled analysis of ITH and TMB show that mPFS and ORR in the TMB-H+ITH-L group was significantly better than that in the TMB-H+ITH-H, TMB-L+ITH-H, TMB-L+ITH-L group (P = 0.0099, P = 0.0121). A subgroup analysis of lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) found that ITH-L was associated with better mPFS (ITH-L versus ITH-H = 8.71 versus 2.17, HR = 3.42, 95%CI 0.96−12.23, P = 0.0460) in LUSC, and no significantly mPFS benefit was found in the ITH-L group in LUAD (ITH-L versus ITH-H = 7.98 versus 5.09, HR = 1.69, 95%CI 0.88−3.26, P = 0.1100). There was no significant difference in TMB levels between ITH-L and ITH-H groups (13.8 Muts/Mb versus 14.2 Muts/Mb, P = 0.2139). Conclusions: ITH may serve as a positive predictor of immunotherapy in patients with advanced NSCLC, especially in LUSC and their clinical value warrants further investigation.

Following an in vitro bioactivity‐guided fractionation procedure, 14 compounds including eight flavonoids and six phenylpropanoids were isolated and identified from the AcOEt fraction of Clinopodium chinense ( Benth .) O. Kuntze . All constituents were tested for α ‐glucosidase and high glucose‐induced injury in human umbilical vein endothelial cells ( HUVEC s) inhibitory activities. All constituents exhibited varying degrees α ‐glucosidase inhibitory activity and protective activity on HUVEC s. Among them, luteolin ( 2 ), eriodictyol ( 5 ), ethyl rosmarinate ( 13 ), and clinopodic acids B ( 14 ) were proved to be potent α ‐glucosidase inhibitors with IC 50 value ranging from 0.6 to 2.0 μ m . Additionally, luteolin ( 2 ), naringenin ( 4 ), eriodictyol ( 5 ), ethyl (2 R )‐3‐(3, 4‐dihydroxyphenyl)‐2‐hydroxypropanate ( 9 ), caffeic acid ( 11 ), ethyl rosmarinate ( 13 ), and clinopodic acids B ( 14 ) significantly ameliorate HUVEC s injury induced by high glucose with an approximate EC 50 value of 3 – 36 μ m . These results suggest that the 14 bioactive constituents were responsible for hypoglycemic and protective vascular endothelium effect of C. chinense ( Benth .) O. Kuntze and their structure–activity relationship was also analyzed briefly. Eriodictyol, luteolin, ethyl rosmarinate, and clinopodic acids B were the potential lead compounds of antidiabetic drugs.

Endothelial dysfunction has been observed in diabetes mellitus, which is related to hyperglycemia. Previous studies have shown that incubation of human umbilical vein endothelial cells (HUVECs) in high glucose causes a significant increase in apoptosis, possibly related to an increase in oxidative stress. The present study aims at investigating the protective effects of Clinopodium chinense extract (CCE) on high glucose-induced apoptosis and the underlying mechanisms. We show that treatment of CCE significantly increased cell viability and the activity of superoxide dismutase, reduced oxidative stress, and protected HUVECs against apoptosis induced by high glucose. Furthermore, our results demonstrate that the antiapoptotic effect of CCE was associated with the reduction of Bax expression and caspase-3 activity and the elevation of Bcl-2 expression. The decrease in caspase-3 activity and caused by CCE were blocked by LY294002, a specific inhibitor of phosphatidylinositol 3-kinase, the upstream kinase of Akt. Moreover, treatment of CCE on HUVECs resulted in dose-dependenent phosphorylation of Akt and inhibited the activation of NF-kappa B. Taken together, our results indicate that CCE could play an important role in protecting endothelial cell by inhibiting oxidative stress and modulating cell apoptosis signaling via the pAkt/NF-kappa B/Bax/Bcl-2/caspase-3 pathway, suggesting that it might be useful for the treatment of endothelial dysfunction-associated diseases such as vascular complications of diabetes mellitus.