Integrated analyses reveal CST7 and DUSP5 regulate Th2 cells differentiation to promote chronic HBV infection.

Chronicity of HBV infection is a complex process influenced by both viral and host factors. Understanding the complex interplay between HBV and cellular immunity is critical. In this study, we used bulk expression datasets for CHB liver tissue from GSE83148 and GSE84044, and scRNA-seq data of CHB liver samples from GSE182159 to find critical genes and immune cells accounted for CHB. We first identified DEGs closely associated with CHB by WGCNA and these genes were intricately linked to differentiation of Th2 cells, which were significantly higher in CHB and positively associated with ALT, AST, HBV-DNA, Scheuer grade and Scheuer stage. Among these DEGs, CST7 and DUSP5 highly expressed in CHB and positively associated with ALT, AST, HBV-DNA, Scheuer grade and Scheuer stage. Moreover, through scRNA-seq, we also found that CST7 and DUSP5 upregulated in Th2 cells and regulated differentiation of naive CD4+ T cells to Th2 cells. Finally, in-vitro studies also showed that HBV infection could significantly up-regulate DUSP5 and CST7 expression. This research strongly revealed that HBV could up-regulate CST7 and DUSP5 to drive differentiation of naive CD4+ T cells into Th2 cells and contribute to CHB, which may pave the way for immunotherapeutic interventions.

Overexpression of ß-Adrenergic Receptors and the Suppressive Effect of ß2-Adrenergic Receptor Blockade in Oral Squamous Cell Carcinoma.

PURPOSE: The purpose of this project was to investigate the expression of ß-adrenergic receptors in oral squamous cell carcinoma (OSCC) and the tumor suppressive activity of ß2-adrenergic receptor (ß2-AR) blockade. MATERIALS AND METHODS: Samples of 15 normal oral mucosal epithelial tissues, 60 surgically resected OSCC tissues, and 60 adjacent para-carcinoma tissues were collected. The expression of ß1-adrenergic receptor and ß2-AR was detected by real-time quantitative polymerase chain reaction and the Western blot test. SCC9 and Cal27 cell lines and primary OSCC cells also were included and treated with ICI-118,551 (MedChemExpress, Monmouth Junction, NJ), a selective ß2-AR blocker. In addition, the Cal27 cell line was treated with propranolol (a nonselective ß-adrenergic receptor blocker) to verify the suppressive effect of ß2-AR blockade. For in vivo assays, Cal27 cells were subcutaneously injected in the tongue flank of nude mice. ICI-118,551 was orally administered to the mice in the treatment group daily. High-throughput sequencing was used to screen for changes in gene expression. RESULTS: Real-time quantitative polymerase chain reaction and the Western blot test both showed that ß1-adrenergic receptor and ß2-AR were overexpressed in OSCC tissues and cells. A relationship was found between ß2-AR and a more advanced clinical stage, as well as preoperative lymphatic metastasis. After treatment with ICI-118,551 or propranolol, the capacities for proliferation, invasion, and metastasis of OSCC cells were significantly inhibited. Tumor size was significantly different between the ICI-118,551 and control groups. The survival time in the ICI-118,551 group also was prolonged significantly. Moreover, high-throughput sequencing identified 19 affected signaling pathways, including mitogen-activated protein kinase and PI3K-Akt. We confirmed a significant change to the expression of several genes closely related to the progression of cancer. CONCLUSION: This study showed that ß2-AR is related to a more advanced clinical stage and preoperative lymphatic metastasis. Additionally, a ß2-AR blocker has a significant suppressive effect in OSCC.

Characterisation of a subpopulation of CD133+ cancer stem cells from Chinese patients with oral squamous cell carcinoma.

Cancer stem cells (CSCs) play a critical role in cancer development and growth. The aim of this study was to identify and isolate CSCs from populations of primary oral squamous cell carcinoma (OSCC) cells, which were obtained from OSCC specimens and identified by cell morphology and immunohistochemical staining for keratin. CD133+ cells detected by flow cytometry comprised 0.41 ± 0.06% of primary OSCC cells and were isolated from primary OSCC cell populations using magnetic-activated cell sorting, revealing that 93.39% of high-purity CD133+ cells were in the G0/G1 phase of the cell cycle. Additionally, the growth rate of CD133+ cells was higher than that of CD133- cells, and in vivo tumourigenesis experiments showed that the tumourigenic ability of CD133+ cells was markedly stronger than that of CD133- cells. Moreover, CD133+ cells showed increased chemotherapeutic resistance to cisplatin and higher self-renewal ability according to sphere-formation assay, as well as higher mRNA levels of stemness-associated genes, including NANOG, SOX2, ALDH1A1, and OCT4. These results indicated that OSCC cells, which share certain characteristics of CSCs, harbour CD133+ cells potentially responsible for OSCC aggressiveness, suggesting CD133 as a potential prognostic marker and therapeutic target.

Expression of beta adrenergic receptor in oral squamous cell carcinoma and its significance to the prognosis.

The aim of this study was to detect the expression of ß-AR (Beta Adregenic Receptor) in Oral squamous cell carcinoma (OSCC), para-cancerous and normal oral mucosa and to investigate the relationship between the expression intensity and the characteristics and prognosis of oral cancer. 100 cases of OSCC were collected; 20 cases of paraneoplastic tissues and 10 cases of normal oral mucosa were taken as control. The expression of ß-AR was detected by immunohistochemical method and the average optical density determination using Image J software. Finally, the difference of ß-AR expression and the correlation with the clinicopathological factors were analyzed statistically. The expression of ß-AR in OSCC was higher than that in paracarcinoma and normal mucosa (P<0.01). The expression intensity of ß1, ß2-AR in preoperative lymph node metastasis group was higher than that in patients without lymph node metastasis (P<0.01). The expression intensity of ß3-AR was not related to pathological grade and tumor size (P>0.05). ß1 and ß2-AR in early stage of OSCC were higher than those in early stage (P<0.05). Lymph node metastasis, recurrence, TNM clinical stage, and the expression intensity of ß1-AR all had an effect on the cumulative survival rate. All the ß1, 2, 3-AR were expressed in OSCC. ß1 and ß2-AR were involved in lymphatic metastasis and had influence on clinical staging. Metastasis, recurrence, TNM stage and expression of ß1-AR had an effect on the cumulative survival rate of tumor. The expression of ß3-AR in OSCC was not associated with the pathological grades and tumor growth.