ABSTRACT
Objective To investigate the effects of microRNA497 (miR-497) on the metastasis of gastric cancer and its possible molecular mechanism. Methods SGC-7901 gastric cancer parent cells were cultured in an ultra-low adhesion environment, and the anoikis resistance model of SGC-7901 cells was created after re-adhesion. Clone formation assay, flow cytometry, TranswellTM test and scratch healing test were used to detect the differences of biological behavior compared with their parent cells. Fluorescence quantitative PCR was performed to detect the expression of miR-497. Western blot analysis was used to detect the changes of key proteins of Wnt/β-catenin signaling pathway and epithelial mesenchymal transformation (EMT) related proteins such as vimentin and E-cadherin. Parent cells and anoikis resistant SGC-7901 cells were transfected with miR-497 inhibitor or miR-497 mimic, and CCK-8 assay was used to detect the proliferation activity. TranswellTM invasion assay was performed to detect the invasion ability of cells. TranswellTM migration test and scratch healing assay was used to determine the migration ability. Western blot analysis was used to detect the expressions of Wnt1, β-catenin, vimentin and E-cadherin. By transfecting miR-497 mimic into the anoikis resistance SGC-7901 cells and inoculating them subcutaneously in nude mice, the changes in the volume and mass of tumor tissues were measured and recorded. Western blot analysis was used to determine the expressions of Wnt1, β-catenin, vimentin and E-cadherin of tumor tissues. Results Compared with the parent cells, the anoikis resistance SGC-7901 gastric cancer cells had faster proliferation rate, stronger colony formation, lower apoptosis rate, stronger invasion and migration ability. The expression of miR-497 was significantly decreased. After down-regulation of miR-497, the proliferation ability, invasion and migration ability were significantly enhanced. The expressions of Wnt1, β-catenin and vimentin increased significantly, while E-cadherin decreased notably. The results of up-regulation miR-497 were the opposite. The tumor growth rate, tumor volume and mass of miR-497 overexpression group were significantly lower than those of control group. The expressions of Wnt1, β-catenin and vimentin decreased significantly, while the expression of E-cadherin increased significantly. Conclusion The expression of miR-497 is low in the anoikis resistance SGC-7901 cells. miR-497 can inhibit the growth and metastasis of gastric cancer cells by blocking Wnt/β-catenin signaling pathway and EMT.
Subject(s)
Animals , Mice , Humans , beta Catenin/metabolism , MicroRNAs/metabolism , Vimentin/metabolism , Stomach Neoplasms/pathology , Anoikis/genetics , Wnt Signaling Pathway/genetics , Mice, Nude , Cell Proliferation/genetics , Cadherins/genetics , Cell Line, Tumor , Epithelial-Mesenchymal Transition/genetics , Cell Movement/geneticsABSTRACT
@#[Abstract] Objective: To investigate the effect of polyadenosine diphosphate ribose polymerase 1 (PARP1) on the proliferation and 5-FU resistance of gastric cancer cells and its potential mechanism. Methods: The tumor tissues and corresponding paracancerous tissues of 72 patients with gastric cancer who were treated in Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from May 2018 to December 2019 were collected. AGS cells were transfected with siFUT8 to knock down FUT8 gene expression. qPCR and immunohistochemistry were used to detect the expression of PARP1 in gastric cancer and adjacent tissues. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis and colony formation of AGS cells. MTT assay was used to detect the effect of AG14361 on the 5-FU sensitivity of gastric cancer cells. The overall distribution of differential genes in AGS cells treated with AG14361 was analyzed by mRNA sequencing, and related signaling pathways were analyzed by KEGG enrichment. qPCR and WB were used to detect the expression of α-1,6-fucosyltransferase (FUT8) in AGS cells and the interference effect of FUT8. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis, and colony formation of AGS cells disturbed by siFUT8. Results: Compared with paracancer tissues, PARP1 expression was significantly increased in gastric cancer tissues (P<0.001). AG14361 can significantly inhibit the proliferation and colony formation of AGS cells, thus promoting the apoptosis of AGS cells (all P<0.01). AG14361 treatment reduced the IC50 of 5-FU against gastric cancer cells, especially against AGS cells, with IC50 decreased by more than 60%. mRNA sequencing results showed that FUT8 was a key glycosyltransferase of AG14361 in inhibiting the proliferation of AGS cells (P<0.05). Compared with the siNC group, treatment of AG14361 with IC50 significantly reversed the promotion of AGS cells proliferation caused by inerference with FUT8, promoted apoptosis and BAX protein expression, decreased Bcl2 protein expression and inhibited the increase in AGS cell colony formation caused by interference with FUT8 (all P<0.01). Conclusion: PARP1 can promote malignant transformation of gastric cancer cells by regulating N-glycosyltransferase FUT8. AG14361 can enhance the chemotherapy sensitivity of 5-FU, and PARP1 may become a potential target for gastric cancer treatment.
ABSTRACT
Objective@#To make a preliminery research of comobidity in oral squamous cell carcinoma (OSCC) patients who resides in Beijing area and investigate whether comorbidity affect the surviving rate independently. Compare the similarities and differences between Chinese and foreign OSCC patients.@*Methods@#The medical records of 313 patients who undertaken operation in Peking University Stomatology School from January 2007 to Delember 2009 were retrospectively reviewed. Adult comorbidity evaluation-27 Chinese edition index was used to estimate the comorbidity severity. COX proportional hazards model was used to analyze whether the TNM stage, comobidity, age and gender affected 5-year survival rate.@*Results@#TNM stage and comorbidity have a significant impact on survival rate, the postoperative survival rate decreased significantly with the increasing level of TNM staging and the complexity of comorbidity disease. In this study, the proportion of patients with none, mild, moderate and severe comorbidity diseases was 24%, 48%, 18% and 10%. The five-year survival rates of patients with moderate and severe comorbidity disease were 50% (29/58) and 13% (4/30) respectively.@*Conclusions@#The comorbidity disease information can help assess the overall health of OSCC patients, and it is recommended to improve the clinical staging and overall evaluation of oral cancer patients with comorbidity disease information.
ABSTRACT
Objective: To investigate the clinical features in patients with respiratory disease and/or hypoxia related severe pulmonary hypertension. Methods: Our research included in 2 groups: Hypoxia related pulmonary hypertension group, the patients with respiratory disease and/or hypoxia combining severe pulmonary hypertension, n=31 and Severe idiopathic pulmonary hypertension group, n=41. The diagnosis was confirmed by right heart catheterization; the patients treated in our hospital from 2016-01 to 2017-01 were consecutively enrolled and studied. Clinical features and treatment were compared between 2 groups. Results: Compared with Severe idiopathic pulmonary hypertension group, the patients' mean age, BMI and blood pressure were higher in Hypoxia related pulmonary hypertension group, while the majority clinical features were similar between 2 groups. In Hypoxia related pulmonary hypertension group, more patients had obstructive sleep apnea-hypopnea syndrome (OSAHS) with the higher AHI and the lower mean blood oxygen pressure at night, increased diameters of left atria and ventricle, elevated NT-proBNP level and reduced 6 minutes walking distance; some patients tried target drug therapy, calcium channel blocker therapy or continuous positive airway pressure therapy. Conclusion: Respiratory disease especially OSAHS should be screened from the patients with severe pulmonary hypertension. In addition to basic respiratory and hypoxia medication, further investigation is needed to confirm whether the prognosis could be improved by calcium channel blocker therapy and target drug therapy.
ABSTRACT
BACKGROUND: Exosomes have the function of some mesenchymal stem cells. Understanding the substance composition that plays a representative role in mesenchymal stem cell exosomes will provide clues for further exploration of synthetic exosome analogues. OBJECTIVE: To investigate the difference of microRNA expression profiles in exosomes derived from passage 2 and 5 human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS: Exosomes in the supernatant of passage 2 and 5 hUC-MSCs were extracted by ultra-high speed centrifugation. The established library was sequenced by using high-throughput sequencing technology. Then we analyzed the sequence results so as to understand the microRNA expression between different groups, and finally did a cluster analysis. RESULTS AND CONCLUSION: 427 657 kinds of microRNAs were detected in the exosomes from passage 2 hUC-MSCs, accounting for 68.93% of the total microRNAs detected; and 119283 microRNAs were detected in the exosomes from passage 5 hUC-MSCs, accounting for 19.22% of the total microRNAs detected. There were 73 526 microRNAs shared between the exosomes from passage 2 and passage 5 hUC-MSCs, accounting for 11.85% of the total microRNAs detected. Bioinformatics analysis (cluster analysis) results showed that these miRNAs were likely to be involved in 161 biological processes, including cell repair, immune and anti-aging. The microRNAs in exosomes from passage 2 to passage 5 hUC-MSCs were largely different. Partial miRNAs exhibited significantly reduced copy numbers. The top five microRNAs with a higher amount, including has-miR-146a-5p, has-miR-191-5p, has-miR-493-3p, has-miR-423-5p, and has-miR-134-5p, have the potential to be the component of synthetic exosome analogues.
ABSTRACT
Objectives: To explore weather oxygen uptake efficiency slope (OUES) may predict the prognosis in patients with idiopathic pulmonary arterial hypertension (IPAH). Methods: The consecutive newly diagnosed IPAH patients in our hospital from 2010-11 to 2015-06 were prospectively enrolled and regular follow-up study was conducted to record cardiovascular events (death and lung transplantation). Kaplan–Meier curve, uni- and multivariate Cox regression analysis were performed to assess the survival rate in relevant patients. Results: A total of 210 IPAH patients at the mean age of (32±10) years were finished cardiopulmonary exercise test (CPET) and received regular follow-up study including 159 female. There were 31 patients died and 1 received lung transplantation over 41 months follow-up period. OUES was positively related to peak oxygen uptake (VO2)/body weight (r=0.71, P0.52 L/(min?m2) (41.9% vs 89.8%), P<0.0001.Conclusion: OUES as a submaximal CPET parameter may well predict the prognosis in IPAH patients.
ABSTRACT
Objective: To observe clinical features in pulmonary hypertension (PH) patients combining obstructive sleep apnea (OSA). Methods: Sleep apnea monitoring was conducted in 65 PH patients with right cardiac catheterization in our hospital from 2016-04 to 2016-08. General clinical tests and the parameters of sleep respiration and right cardiac catheterization were recorded. OSA was diagnosed by apnea hypopnea index (AHI)≥5, different parameters were compared between PH patients with and without OSA. Results: The average patients' age was (41.98±15.26) years including 72.31% (47/65) female, 26 (40%) patients combining OSA with the mean AHI at (18.12±13.40). Compared to those without OSA, PH with OSA patients had the elder age, more male and higher proportions of chronic thromboembolic PH, lung diseases or hypoxia; increased AHI, apnea index (AI), obstructive AI (oAI); more patients with nocturnal hypoxia>10% and SaO2<90%, increased BMI, more NYHA Ⅲ and elevated NT-proBNP; while decreased mean oxygen saturation, minimum oxygen saturation, PaO2, 6 minute walk distance and cardiac index, all P<0.05. Drug therapy was similar between 2 groups. Conclusion: Nocturnal hypoxia and OSA were common in PH patients, elder age and male gender were the risk factors for PH combining OSA; the patients had lower partial pressure of oxygen especially at night, longer time of hypoxia and severer cardiac function damage. It is necessary to conduct sleep apnea monitoring to alert sleep apnea and hypoxia in relevant patients.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of emodin on the contraction of jejunum smooth muscle and its underlying mechanisms.</p><p><b>METHODS</b>Rats were randomly divided into 7 groups (n = 6): control group, emodin group (1, 5, 10, 20 micromol/L), propranolol (PRO) plus emodin group, glibenclamide (GLI) plus emodin group, NG-Nitro-L-arginine Methyl Ester (L-NAME) plus emodin group, calcium free control group and calcium free emodin group. The rats were sacrificed by cervical dislocation and the small intestine was isolated. The jejunum segment specimens were mounted on an Organ Bath System with a tension transducer. The effect of emodin on contraction of jejunum smooth muscle was measured by BL-420E+ biological signal processing system and the amplitude (AM), tension (TE) and frequency (FR) of contraction were determined.</p><p><b>RESULTS</b>(1) Emodin inhibited the tension and amplitude of jejunum smooth muscle contraction in a dose-dependent manner (P < 0.05, P < 0.01) while the frequency was not obviously influenced. (2) PRO (P < 0.05) or GLI (P < 0.01) partly abolished the inhibitory effect of emodin on jejunum smooth muscle. (3) L-NAME had no obvious effect on the inhibitory effect of emodin. (4) Emodin attenuated the contraction of jejunum smooth muscle induced by calcium chloride application into calcium free K-H solution (P < 0.01).</p><p><b>CONCLUSION</b>Emodin obviously inhibits the amplitude and tension, while has no influence on the frequency of jejunum smooth muscle contraction in rats. Activation of beta adrenergic receptor, open of ATP sensitive potassium channels, and inhibition of the extracellular calcium influx through calcium channels of smooth muscle cell membrane might be involved in the process.</p>