Protective effects of salvianolic acid B on intestinal ischemia/reperfusion injury in rats by regulating the AhR/IL-22/STAT6 axis.

PURPOSE: Intestinal ischemia/reperfusion (I/R) injury (IIRI) is associated with high morbidity and mortality. Salvianolic acid B (Sal-B) could exert neuroprotective effects on reperfusion injury after cerebral vascular occlusion, but its effect on IIRI remains unclear. This study set out to investigate the protective effects of Sal-B on IIRI in rats. METHODS: The rat IIRI model was established by occluding the superior mesenteric artery and reperfusion, and they were pretreated with Sal-B and aryl hydrocarbon receptor (AhR) antagonist CH-223191 before surgery. Pathological changes in rat ileum, IIRI degree, and intestinal cell apoptosis were evaluated through hematoxylin-eosin staining, Chiu's score scale, and TUNEL staining, together with the determination of caspase-3, AhR protein level in the nucleus, and STAT6 phosphorylation by Western blotting. The levels of inflammatory cytokines (IL-1ß/IL-6/TNF-α) and IL-22 were determined by ELISA and RT-qPCR. The contents of superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) in intestinal tissues were determined by spectrophotometry. RESULTS: Sal-B alleviated IIRI in rats, evidenced by slight villi shedding and villi edema, reduced Chiu's score, and diminished the number of TUNEL-positive cells and caspase-3 expression. SAL-B alleviated inflammation and oxidative stress (OS) responses induced by IIRI. Sal-B promoted IL-22 secretion by activating AhR in intestinal tissue after IIRI. Inhibition of AhR activation partially reversed the protective effect of Sal-B on IIRI. Sal-B promoted STAT6 phosphorylation by activating the AhR/IL-22 axis. CONCLUSION: Sal-B plays a protective role against IIRI in rats by activating the AhR/IL-22/STAT6 axis, which may be achieved by reducing the intestinal inflammatory response and OS responses.

Audiological characteristics and effect of endoscopic surgery for simple congenital malformation of ossicular chain.

OBJECTIVE: To analyze the audiological characteristics and surgical results in patients undergoing surgery for simple congenital ossicular chain malformation, and the effect of endoscopic surgery. METHODS: A retrospective review was performed on 86 patients who underwent surgery for the congenital malformation of the ossicular chain. Clinical characteristics and audiometric data were analyzed. Fifty-eight patients had detailed postoperative data, and the preoperative and postoperative audiometric results were compared. The subjects were further divided into endoscopic and microscopic groups, and their surgical effects were examined. RESULTS: The preoperative audiometry results in the low-frequency group were worse than those in the high-frequency group (P < 0.05). A postoperative air-bone gap closure to 20 dB or less was achieved in 73.33% of the 60 ears of patients postoperatively. The postoperative air conduction and air-bone gap were significantly better than the preoperative ones (P < 0.05), and the improvement effect was the best in class III patients (P < 0.05). Postoperative hearing had no significant differences between the endoscopic and microscopic groups. However, endoscopic surgery also was more advantageous in terms of operating time (P < 0.05). CONCLUSIONS: Preoperative pure tone audiometric results showed moderate or moderate-severe hearing loss, especially in the low-frequency area. The reconstruction of the auditory ossicle chain can achieve satisfactory results, especially in class III patients. Endoscopic and microscopic surgery in the treatment of simple congenital ossicular chain malformations can effectively improve postoperative hearing.

Outcomes of Recurrent Nasopharyngeal Carcinoma Patients Treated With Salvage Surgery: A Meta-Analysis.

OBJECTIVE: To assess the efficacy of treatment outcomes of salvage surgery for recurrent nasopharyngeal carcinoma (rNPC). METHODS: We conducted a detailed search of the literatures in biomedical databases published from January 1990 to December 2020. The main research features and results of interest were retrieved from the articles that met the selection criteria for meta-analysis. RESULTS: A total of 21 articles with 778 patients were included, 17 of which met the meta-analysis inclusion criteria. The pooled 2-year overall survival (OS), 5-year OS, and 2-year disease-free survival (DFS) were 71%, 50% and 61%, respectively. Subgroup analysis was conducted with postoperative adjuvant therapy. The pooled 2-year OS, 5-year OS and 2-year DFS of the postoperative adjuvant therapy group compared with the surgery alone group were 69% vs 72%, 44% vs 56%, and 77% vs 54%, respectively. Univariate and multivariate analyses were performed on 178 patients with detailed individual postoperative survival data in 10 articles. On multivariate analysis, recurrent T (RT) stage and adjuvant therapy were independent predictors of outcomes. CONCLUSIONS: This meta-analysis indicated that recurrent NPC patients can obtain survival benefits from salvage surgery. Accurately assessing the RT stage of the tumor and choosing the appropriate surgical method are important to the success of the surgery. Although the prognostic factors influencing outcome have been studied, conclusive data on the survival benefits are still lacking. Random controlled trials (RCTs) to compare surgery alone and postoperative adjuvant therapy are needed in patients with positive margin status after salvage surgery.

MicroRNA-328-3p facilitates the progression of gastric cancer via KEAP1/NRF2 axis.

Gastric cancer is a common lethal malignancy and causes great cancer-related mortality worldwide. MicroRNA (miR)-328-3p is implicated in the progression of various human cancers; however, its role and mechanism in the progression of gastric cancer remain unclear.Human gastric cancer cells were incubated with miR-328-3p mimic, inhibitor or the matched negative control. Cell viability, colony formation, migrative and invasive capacity, cell apoptosis and oxidative stress were measured. To clarify the involvement of nuclear factor-E2-related factor 2 (NRF2) and kelch-like ECH-associated protein 1 (KEAP1), small interfering RNA was used. miR-328-3p was upregulated in human gastric cancer cells and tissues, and its level positively correlated with the progression of gastric cancer. miR-328-3p promoted cell viability, colony formation, migration and invasion, thereby facilitating the progression of gastric cancer. miR-328-3p mimic reduced, while miR-328-3p inhibitor increased apoptosis and oxidative stress of human gastric cancer cells. Mechanistically, miR-328-3p upregulated NRF2 via targeting KEAP1to attenuate excessive free radical production and cell apoptosis. miR-328-3p functions as an oncogenic gene and inhibiting miR-328-3p may help to develop novel therapeutic strategies of human gastric cancer.

Sophoridine exerts tumor-suppressive activities via promoting ESRRG-mediated ß-catenin degradation in gastric cancer.

BACKGROUND: As a natural alkaloid product isolated from Sophora alopecuroides. L, Sophoridine reshapes gastric cancer immune microenvironment via inhibiting chemotaxis and M2 polarization of tumor-associated macrophages (TAMs). However, the exact effects and underlying mechanism of Sophoridine on gastric cancer cells remains poorly known. METHODS: The potential anti-tumor effects of Sophoridine on gastric cancer cell lines, including AGS and SGC7901 cells, were detected by CCK-8, EDU and colony forming assay, immunofluorescence, transwell assay, and flow cytometry. Molecular mechanisms of Sophoridine were investigated by siRNA transfection, nuclear/cytoplasmic extraction and western blot. The synergistic effects of Sophoridine with cisplatin on gastric cancer cells were further investigated in in vitro functional studies. RESULTS: Sophoridine exhibited potent tumor-suppressive activities in gastric cancer cells, including inhibition of proliferation, colony formulation, migration and invasion, as well as induction of apoptosis. In addition, we further showed that Sophoridine induced G2/M cell cycle arrest via inhibiting double-stranded DNA breaks repair and enhanced the efficacy of cisplatin in gastric cancer cells. Molecular studies further revealed that Sophoridine promoted ß-catenin degradation by enhancing Estrogen-related receptor gamma (ESRRG) expression, but not depended on ubiquitination-proteasome pathway, either TRIM33-mediated (GSK3ß-independent) or altered GSK3ß activity, and thus exerted potent tumor-suppressive activities. CONCLUSION: Sophoridine depends on targeting ESRRG/ß-catenin pathway to exert tumor-suppressive activities in gastric cancer cells and enhances the anti-tumor effect of cisplatin. Our study provided the promising preclinical anti-tumor evidence for the potential application of Sophoridine against gastric cancer.

The positive feedback loop FOXO3/CASC11/miR-498 promotes the tumorigenesis of non-small cell lung cancer.

An increasing number of studies have indicated that long noncoding RNAs (lncRNAs) are involved in the regulation of non-small-cell lung cancer (NSCLC). Nevertheless, there are still numerous undiscovered mechanisms underlying this molecular regulation. Here, the results illustrated that CASC11 is overexpressed in NSCLC tumor tissues and cell lines, which is closely related to the clinical features of NSCLC and poor survival. In functional experiments, CASC11 was shown to promote proliferation and cycle progression and enhance NSCLC tumorigenesis. In mechanical investigations, CASC11 was shown to target the miR-498/FOXO3 axis via a canonical competing endogenous RNA (ceRNA). In return, the transcription factor FOXO3 targets the CASC11 promoter region, thereby accelerating its transcription. Our findings demonstrate a crucial role for CASC11 as an oncogene in promoting NSCLC. These results reveal that CASC11 might be a potential therapeutic target for NSCLC.

MnTE-2-PyP Attenuates TGF-ß-Induced Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Inhibiting the Smad2/3 Signaling Pathway.

BACKGROUND: As a key step in enhancing cancer cell invasion and metastasis, epithelial-mesenchymal transition (EMT) plays an important role in colorectal cancer progression. EMT is triggered by a variety of signaling pathways, among which the transforming growth factor ß (TGF-ß) signaling pathway has been implicated as a primary inducer. Accumulating evidence demonstrates that MnTE-2-PyP (chemical name: manganese(III) meso-tetrakis-(N-ethylpyridinium-2-yl), a superoxide dismutase (SOD) mimetic, inhibits TGF-ß signaling; however, its ability to inhibit TGF-ß-induced EMT in colorectal cancer has not yet been explored. METHODS: To verify our hypothesis that MnTE-2-PyP attenuates TGF-ß-induced EMT, human colorectal cancer cells were treated with TGF-ß in the presence or absence of MnTE-2-PyP. Cells were analyzed by several techniques including western blotting, real-time quantitative PCR, transwell assay, and wound healing assay. RESULTS: MnTE-2-PyP reverses cell phenotypes induced by TGF-ß in colon cancer cells. MnTE-2-PyP treatment significantly reduced the expression of mesenchymal markers but maintained epithelial marker expression. Mechanistically, MnTE-2-PyP suppressed the phosphorylated Smad2/3 protein levels induced by TGF-ß in SW480 cells, but MnTE-2-PyP failed to suppress TGF-ß-induced Slug and Snail expression in colorectal cells. Furthermore, MnTE-2-PyP effectively suppressed TGF-ß-mediated cell migration and invasion and the expression of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) in colorectal cells. CONCLUSION: Taken together, we provide an in-depth mechanism by which MnTE-2-PyP inhibits colorectal cancer progression, supporting an important role for MnTE-2-PyP as an effective and innovative antitumor agent to enhance treatment outcomes in colorectal cancer.

SOX2 inhibits cell proliferation and metastasis, promotes apoptotic by downregulating CCND1 and PARP in gastric cancer.

Inconsistent results of Sex-determining region Y-box2 (SOX2) expression have been reported in gastric cancer (GC) before. Our recent studies showed that SOX2 was significantly downregulated in GC cells compared with GES-1 at both mRNA and protein level. Transfected with pcDNA3.1-SOX2 resulted in enforced expression of SOX2 at mRNA and protein levels compared with NC group in undifferentiated cell lines including HGC27 and BGC823. MTT assay showed that exogenous expressed SOX2 suppressed cell proliferation. FC analysis revealed that SOX2-overexpressing cells exhibited cell-cycle arrest and apoptosis. Transwell assay showed the anti-metastatic effect of SOX2 in GC cells. The subsequent results suggested CCND1 and PARP were downregulated in SOX2 overexpressed GC cells, and were responsible for the SOX2-induced anticancer effects. Thus, SOX2 proved to be an expected biomarker in GC diagnosis.

Constitutive activation of STAT3 and cyclin D1 overexpression contribute to proliferation, migration and invasion in gastric cancer cells.

Signal transducer and activator of transcription-3 (STAT3) is closely associated with tumorigenesis and is activated in tumor cells of a variety of cancers. The raised expression of cyclin D1 (CCND1) by activated STAT3 has been verified in some cancers. However, the relationship between STAT3 and CCND1 has yet to be studied in gastric cancer (GC) cells. In the present study, we found that STAT3 was constitutively activated in several GC cells together with overexpressed CCND1. In addition, IL-6 treatment enhanced the expression level of p-STAT3 and CCND1, accelerating the cell cycle progress and transferring from G1 to S phase. The increased proliferation, migration and invasion were also demonstrated by the treatment of IL-6 in HGC-27 and BGC-823 cells. While AG490 treatment, a Janus Kinase (JAK) inhibitor, showed the opposite effect. Therefore, our research demonstrated the positive correlation between p-STAT3 and CCND1 in GC cells. The constitutive activation of STAT3 and CCND1 overexpression accounted for the proliferation, migration and invasion in GC cells.

Targeting Rad50 sensitizes human nasopharyngeal carcinoma cells to radiotherapy.

BACKGROUND: The Mre11-Rad50-Nbs1 (MRN) complex is well known for its crucial role in initiating DNA double strand breaks (DSBs) repair pathways to resistant irradiation (IR) injury and thus facilitating radioresistance which severely reduces radiocurability of nasopharyngeal cancer (NPC). Targeting native cellular MRN function would sensitize NPC cells to IR. METHODS: A recombinant adenovirus containing a mutant Rad50 gene (Ad-RAD50) expressing Rad50 zinc hook domain but lacking the ATPase domain and the Mre11 interaction domain was constructed to disrupt native cellular MRN functions. The effects of Ad-RAD50 on the MRN functions were assessed in NPC cells lines using western blot, co-immunoprecipitation and confocal microscopy analyses. The increased radiosensitivity of transient Ad-RAD50 to IR was examined in NPC cells, including MTT assay, colony formation. The molecular mechanisms of radiosensitization were confirmed by neutral comet assay and western bolts. Nude mice subcutaneous injection, tumor growth curve and TUNEL assay were used to evaluate tumor regression and apoptosis in vivo. RESULTS: Rad50 is remarkably upregulated in NPC cells after IR, implying the critical role of Rad50 in MRN functions. The transient expression of this mutant Rad50 decreased the levels of native cellular Rad50, Mre11 and Nbs1, weakened the interactions among these proteins, abrogated the G2/M arrest induced by DSBs and reduced the DNA repair ability in NPC cells. A combination of IR and mutant RAD50 therapy produced significant tumor cytotoxicity in vitro, with a corresponding increase in DNA damage, prevented proliferation and cell viability. Furthermore, Ad-RAD50 sensitized NPC cells to IR by causing dramatic tumor regression and inducing apoptosis in vivo. CONCLUSION: Our findings define a novel therapeutic approach to NPC radiosensitization via targeted native cellular Rad50 disruption.

Association between STAT3 polymorphisms and cancer risk: a meta-analysis.

Five polymorphisms, rs2293152, rs4796793, rs12949918, rs6503695, rs744166, in the STAT3 gene have been implicated in susceptibility to cancer, but the results were inconclusive. The aim of this meta-analysis is to investigate the association between the five polymorphisms and cancer risk. All eligible case-control studies published up to March 2015 were identified by searching PubMed, Web of Science, Wanfang, VIP, and CNKI. Effect sizes of odds ratio (OR) and 95 % confidence interval (95 % CI) were calculated by using a fixed- or random-effect model. A total of 15 articles were included. Overall, a significantly decreased risk was found for rs12949918 polymorphism (dominant model: OR = 0.83, 95 % CI: 0.75-0.91, recessive model: OR = 0.77, 95 % CI: 0.68-0.87, TC vs. TT: OR = 0.87, 95 % CI: 0.79-0.96, CC vs. TT: OR = 0.71, 95 % CI: 0.62-0.81), and for rs744166 polymorphism (recessive model: OR = 0.75, 95 % CI: 0.58-0.98; GG vs. AA: OR = 0.68, 95 % CI: 0.51-0.90), while there was no significant association for other three polymorphisms under all genetic models. In subgroup analysis by ethnicity, for rs12949918 polymorphism, similar results were detected among Caucasians, similarly, a significant decreased risk was observed in Asians under dominant and CC vs. TT model; for rs2293152 polymorphism, significant association was detected among Asians under recessive model. This meta-analysis suggests that the STAT3 rs12949918 and rs744166 polymorphisms, but not other three polymorphisms, may be an important protective factor for cancer.

Serine/threonine kinase 15 gene polymorphism and risk of digestive system cancers: A meta-analysis.

Previous studies have reported an association between the two coding polymorphisms (91T>A and 169G>A) of the serine/threonine kinase 15 (STK15) gene and the risk of digestive system cancers; however, the results are inconsistent. In the present study, a meta-analysis was carried out to assess the association between the two STK15 polymorphisms and the risk of digestive system cancers. Relevant studies were identified using PubMed, Web of Science, China National Knowledge Infrastructure, WanFang and VIP databases up to February 18, 2014. The pooled odds ratio (OR) with a 95% confidence interval (CI) was calculated using the fixed or random effects model. A total of 15 case-control studies from 14 publications were included. Of these, 15 studies concerned the 91T>A polymorphism and included 7,619 cases and 7,196 controls and four studies concerned the 161G>A polymorphism and included 826 cases and 713 controls. A significantly increased risk of digestive system cancers was observed for the 91T>A polymorphism (recessive model: OR, 1.19; 95% CI, 1.07-1.31). In subgroup analysis by ethnicity, a significant association was detected in Asian populations (recessive model: OR, 1.21; 95% CI, 1.08-1.36) but not in Caucasian and mixed populations. Stratification by tumor type indicated that the 91T>A polymorphism was associated with an increased risk of esophageal and colorectal cancers under the recessive model (OR, 1.19; 95% CI, 1.03-1.38; and OR, 1.24; 95% CI, 1.04-1.46; respectively); however, no significant association was observed between the 169G>A polymorphism and the risk of digestive system cancers in any of the genetic models. Furthermore, in subgroup analysis by ethnicity, similar results were observed in the Asian and Caucasian populations. The present meta-analysis demonstrated that the STK15 gene 91T>A polymorphism, but not the 169G>A polymorphism, may be a risk factor for digestive system cancers, particularly for esophageal and colorectal cancers.

[The preparation of recombinant adenovirus Ad-Rad50-GFP and detection of the optimal multiplicity of infection in CNE1 transfected hv Ad-Rad50-GFP].

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION: Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.

XPG Asp1104His polymorphism and gastrointestinal cancers risk: a meta-analysis.

Several studies have reported the association between the Asp1104His polymorphism in xeroderma pigmentosum group G (XPG) gene and risk of gastrointestinal cancers. However, the results are inconsistent. This meta-analysis was performed to assess the association between XPG Asp1104His polymorphism and gastrointestinal cancers risk. Relevant studies were identified using PubMed, Web of Science, CNKI, WanFang and VIP databases up to July 22, 2014. The pooled odds ratio (OR) with a 95% confidence interval (CI) was calculated using the fixed- or random effects model. 13 case-control studies from twelve publications with 4275 patients and 5735 controls were included. Overall, a significant association was found between the XPG Asp1104His polymorphism and the risk of gastrointestinal cancers (dominant model: OR = 1.15, 95% CI: 1.05-1.26; His/His vs. Asp/Asp: OR = 1.15, 95% CI: 1.01-1.32). When the analysis was stratified by ethnicity, similar results were observed in Asians under homozygote model; in stratification analysis by cancer type, increased cancer risk was detected in colorectal and hepatocellular carcinoma, but not for other gastrointestinal cancers. Furthermore, in subgroup analysis by source of control, we failed to detect any association among population, hospital and family-based populations. This meta-analysis indicated that the XPG Asp1104His polymorphism may be a risk factor for gastrointestinal cancers, especially of colorectal cancer.

[Radiobiological characteristics and MRN complex expression of human nasopharyngeal carcinoma cell lines].

OBJECTIVE: To study the radiobiological characteristic of human nasopharyngeal carcinoma cell lines CNE1 and CNE2 and the changes in expression MRN (Mre11-Rad50-Nbs1) complex in the cell lines exposed to irradiation. METHODS: CNE1 and CNE2 were irradiated by a linear accelerator. Radiobiological characteristics were detected by colony assay and MTT assay. MRN complex expression were examined by Western blot. RESULTS: Surviving fraction at 2 Gy (SF2), quasi-threshold Dose (Dq), and mean lethal dose (Do) of CNE1 were 0.56, 1.449 Gy and 1.480 Gy; SF2, Dq, and Do of CNE2 were 0.44, 0.776 Gy and 1.685 Gy, respectively. Survival fraction of CNE1 at the day 6 after 4 Gy irradiation was 0.59 and that of CNE2 was 0.79 when compared with control, with the up-regulated expressions of Rad50 in CNE1 and Mre11, Rad50 and Nbs1 in CNE2 (P < 0.05). CONCLUSIONS: CNE1 and CNE2 were sensitive to radiation, but there were radioresistance cells in CNE2. The expressions of some components of MRN complex were up-regulated to repair DNA lesions induced by radiation.