scholarly journals Study on Molecular Mechanism of Benzo (ɑ) pyrene on CMA by HSP90ɑ and HIF-1ɑ

2021 ◽  
Author(s):  
Shasha Zhang ◽  
Tingting Liu ◽  
Qi Chen ◽  
Min Su ◽  
Tuya Bai ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blot ◽  
Nude Mice ◽  
Corn Oil ◽  
A549 Cells ◽  
Tumor Formation ◽  
Control Group ◽  
Transplanted Tumors ◽  
Transplanted Tumor ◽  
Significant Difference

Abstract OBJECTIVE: The effects of benzo (α) pyrene (BaP) on molecular chaperone autophagy (CMA) through heat shock protein 90 (HSP90) and hypoxia inducible factor-1 (HIF-1) were studied by RNA interference and subcutaneous tumor formation technique in nude mice. METHODS: 40 nude mice inoculated with the shHSP90ɑ A549 cell line under the armpits of the forelimbs were divided into 4 groups, 1.80 mg/kg/d BaP-corn oil solution was intragastrically administered for 60 days (except the Control group), and the growth of nude mice and transplanted tumors was recorded curve. The size and morphological changes of tumors were observed by small animal imaging technique and HE staining method. qPCR, Western blot and Immunohistochemistry were used to detect the expression of HSP90ɑ, HSC70 and Lamp-2A. A549 cells were treated with 0.1μmol/L, 1μmol/L and 10μmol/L BaP for 24h, EPO, HIF-1ɑ concentration and HIF-1ɑ protein expression were detected by Elisa and Western blot; A549 cells were treated with 10μmol/L BaP and added HIF-1ɑ inhibitor 24h, qPCR, Western blot and Immunofluorescence method were used to detect the expression of HSP90ɑ, HSC70 and Lamp-2A. RESULTS: BaP can reduce silence HSP90ɑ the weight of nude mice and transplanted tumors (P<0.01); BaP can reduce silence HSP90ɑ the expression of HSP90ɑ, HSC70, Lamp-2A mRNA and protein in transplanted tumor tissues (P<0.05); BaP has no significant difference in the organization structure of silence HSP90ɑ and non-silence HSP90ɑ; BaP can reduce the total number of bioluminescence photons of silence HSP90ɑ transplanted tumors (P<0.01). 10μmol/L BaP can increase the concentration of EPO and HIF-1ɑ and the expression of HIF-1ɑ protein in A549 cells (P<0.05); and after adding HIF-1ɑ inhibitors treat A549 cells, HSP90ɑ, HSC70, Lamp-2A mRNA and protein expression were decreased (P<0.05), and the fluorescence intensity of HSP90ɑ was decreased. CONCLUSIONS: BaP can promote the growth of transplanted tumor in nude mice, while the growth of transplanted tumor in nude mice is inhibited shHSP90ɑ. BaP promotes the occurrence of CMA by promoting the expression of HSP90ɑ and HIF-1ɑ, which are important regulatory genes for BaP to activate CMA.

scholarly journals Study on the Biological Characteristics of CD133+ Cells Interfered by RNA Interference in Gastric Cancer

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Ji-wei Yu ◽  
Shou-lian Wang ◽  
Ju-gang Wu ◽  
Rui-qi Lu ◽  
Xiao-chun Ni ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cancer Cells ◽  
Nude Mice ◽  
Interference Effect ◽  
Tumor Formation ◽  
Biological Characteristics ◽  
Control Group ◽  
Gastric Cancer Cells ◽  
Transplanted Tumor ◽  
Sphere Formation

Background. To detect the changes of biological characteristics in gastric cancer cells interfered by CD133-specific small interfering RNA (siRNA). Methods. First to select the siRNA which has the strongest interference effect among 3 siRNAs (i.e., siRNA1, siRNA2, and siRNA3) in KATO-III cells by RT-PCR and Western blotting assays. Then, CD133+ cells were sorted out from KATO-III cells using an immunomagnetic bead sorting method and transfected with the selected siRNA. Furthermore, the proliferating characteristics, the antichemotherapeutic assessment, Transwell invasion assay, monoclonal sphere formation assay, and subcutaneous transplanted tumor formation assay in nude mice were investigated. Results. siRNA3 showed the strongest interference effect in KATO-III cells. As compared to the uninterfered control group, the CD133+ cells treated by siRNA3 showed significant decreases in the abilities of proliferation, invasion, clone sphere formation, and resistance to antitumour drugs as well as the weight and size of the transplanted tumor, which was nearly similar to that of CD133− cells. Additionally, the protein expression level of the EMT factor E-cadherin increased while those of EMT-related Snail and N-cadherin decreased in CD133+ cells interfered by siRNA3. Conclusion. Inhibition of CD133 gene expression reduces the abilities of gastric cancer cells in proliferation, invasion, clonal sphere formation, and chemoresistance as well as tumor formation in nude mice.

scholarly journals Combination of ω-3 Polyunsaturated Fatty Acids with 5-FU Inhibits SGC7901 Gastric Cancer Cell Growth via Rho-ROCK1 Signaling Pathway in Nude Mice

2021 ◽  
Author(s):  
Jiang Yiyan ◽  
Wang Keke ◽  
Lou Zhefeng ◽  
Hong Dan ◽  
Min Tao
Keyword(s):  
Gastric Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Nude Mice ◽  
Protein Level ◽  
Combination Group ◽  
Control Group ◽  
Mrna Levels ◽  
Caspase 9 ◽  
Gastric Cancer Cells

Abstract Background: Gastric cancer is one of the most common malignancy with high mortality rate in the world. Systemic chemotherapy is thought to be an important treatment. However, due to the unsatisfactory efficiency and obvious side effects, it is urgent to detect new therapy strategy for gastric cancer. This study was aimed to investigate the effects and mechanisms of ω-3 polyunsaturated acids (PUFAs) combined with 5-FU on the growth of gastric cancer cells in nude mice. Methods: BALB/C nude mice were injected subcutaneously with SGC7901 gastric cancer cells to establish a tumor-bearing mouse model. The tumor growth in vivo was observed. Morphological of tumor specimens was observed by HE staining. The mRNA levels of RhoA, RhoC and ROCK1 in tumor tissues were detected by qPCR, and their protein levels were detected by immunofluorescence and Western Blot. Meanwhile, apoptosis –related proteins were also determined by Western Blot.Results: Compared with the NC control group, the tumor volume and weight in ω-3 PUFAs and 5-Fu groups were insignificantly lower, but significantly lower in the combination group. Compared with the abundant blood supply in the NC group, HE staining showed multifocal tumor necrosis in the three intervention groups, and this change was the most prominent in the combination group. And qPCR results showed that the mRNA levels of RhoA in the combination groups were significantly lower than this in the other groups. Immunofluorescence showed that the level of RhoA protein in the three intervention groups decline in varying degrees, especially in the combination group. Western Blot showed that the protein level of RhoA in the three intervention groups were significantly lower than those in the NC control group, especially in the combination group. Meanwhile, the protein level of ROCK1 in both 5-FU group and the combination group were significantly lower, especially in the combination group. Compared with the control group, the levels of Bcl-2 and Caspase-9 decreased in the combination group, the level of cleaved Parp was increased at the same time.Conclusion: ω-3 PUFAs combined with 5-FU may inhibit tumor growth through the Rho/ROCK pathway and promote apoptosis by down-regulating the levels of Bcl-2 and Caspase-9 and induce the increase of cleaved Parp level.

The Effect of Thymol on Renal Toxicity Induced by Mercury Chloride in Rats

2021 ◽  
Author(s):  
Hamid Reza Jamshidi ◽  
Faezeh Taheri
Keyword(s):  
Mercuric Chloride ◽  
Renal Toxicity ◽  
Corn Oil ◽  
Kidney Tissue ◽  
Control Group ◽  
Mercury Chloride ◽  
Protective Effects ◽  
Tissue Samples ◽  
Significant Difference ◽  
Anti Oxidant

Background and Aims: Mercuric chloride is highly toxic once absorbed into the bloodstream, especially the kidneys in which it is collected. Mercury chloride increases hydrogen peroxide and enhances the destruction of protective enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPX), leading to oxidative stress. Besides, thymol has anti-oxidant effects and can increase the activity of SOD and GPX. This study aims to evaluate the efficacy of thymol on mercury chloride-induced toxicity. Materials and Methods: In this study, 30 rats, consisting of 6 groups of 5, were used. Control group receiving a single dose of 0.5 mg/kg mercuric chloride for 15 days, third, fourth, and fifth group received intraperitoneal injection of mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at a dose of 10, 30, 50 mg/kg. The sixth group received mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at 30 mg/kg per day for ten days. Results: Results showed a significant difference in the activity of catalase enzyme in kidney tissue samples test. According to the results of SOD, there is a significant difference between the group of corn oil and the group of mercury chloride and between the group of mercury chloride and the group that receives thymol at a dose of 10, 30, 50 mg/kg (p ≤ 0.05). Conclusions: It can be concluded that mercury chloride-induced kidney toxicity and thymol have anti-oxidant protective effects for SOD and GPX.

Phenolhexyl Isothiocyanate, a New Histone Deacetylase Inhibitor, Can Prevent Tumor Formation and Inhibit Leukemia Cell Growth In Vivo.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4625-4625
Author(s):  
Lulu Lu ◽  
Xudong Ma ◽  
A. Becklemisheva ◽  
J.W. Chaio ◽  
Delong Liu
Keyword(s):  
Cell Growth ◽  
Apoptotic Cell ◽  
Leukemia Cell ◽  
Chemotherapeutic Agents ◽  
Tumor Formation ◽  
Control Group ◽  
Immunodeficient Mice ◽  
Standard Design ◽  
Significant Difference

Abstract We have demonstrated that phenolhexyl isothiocyanate (PHI) induces growth arrest and apoptosis in leukemia cells HL-60 through inhibition of the activity of histone deacetylases (HDAC), the enhancement of histone acetylation and activation of p21. In this study, we examined the effects of PHI on the growth of HL-60 leukemic xenograft in immunodeficient mice. The PHI was given to the mice by gavage. The maximum tolerated dose by the mice was determined following standard design. To determine the in vivo effect of PHI on leukemia growth, a sub-MTD dose was given to the control and the treatment groups with 17 mice in each group after injection of 1.0 x 10e6 HL-60 cells per mouse. There was a significant reduction in the incidence of tumor formation (94.1% control group vs 58.5% PHI group, p=0.004). In addition, there was also a significant difference in the tumor size between the two groups. Determined at autopsy, the mean weight of control tumors was 0.8 g vs. the tumors of the experimental group 0.35g, revealing a significant reduction of 44.4% (P<0.03). There were no detectable toxicity as evaluated by body and organ weight, and necropsy examination. The histology of the tumor showed increased apoptotic cell death. Apoptosis in the tumors was further confirmed by the cleavage of poly ADP-ribose polymerase (PARP), the target of proteolysis of caspases that execute apoptosis with Western blot analyses. The results suggest that PHI can prevent tumor formation and inhibit leukemia cell growth in vivo without significant toxicity that is routinely associated with conventional chemotherapeutic agents.

Lidocaine Inhibits Ovarian Cancer Cell Proliferation and Induces Apoptosis: An In Vitro Study

2020 ◽  
Vol 10 (5) ◽  
pp. 724-729
Author(s):  
Yaping Xu ◽  
Xiaoqin Fang ◽  
Xianjiang Wei
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Western Blot ◽  
Cancer Cell ◽  
Blot Analysis ◽  
Group Treatment ◽  
Western Blot Analysis ◽  
Ovarian Cancer Cell ◽  
Control Group

Objective: The present study aimed to explore the effects and related mechanism of lidocaine on human ovarian cancer cell lines. Methods: Human ovarian cancer cell lines (SKOV3 and ES-2) were treated with different concentrations of lidocaine for different time. We treated SKOV3 and ES-2 cells using lidocaine then used MTT assay and flow cytometry to detect the cell proliferation and cell apoptosis. In addition, we used western blot analysis to explore the protein expression of Bax and Bcl-2 in SKOV3 and ES-2 cells. Western blot analysis and qRT-PCR were performed for the detection of EMT markers (E-cadherin, N-cadherin). The protein expression levels of TRAF3 and p-p65 in SKOV3 and ES-2 cells were determined by Western blot analysis. Results: Compared to the control group, 0.5, 1, 5, and 10 mM of lidocaine significantly inhibited ovarian cancer cell proliferation at different time points, while 0.1 mM of lidocaine had no significant effect. 1, 5 mM of lidocaine induced the cell apoptosis, and observably reduced expression of Bcl-2 protein, but improved Bax expression markedly compared with the control group. Treatment of lidocaine increased E-cadherin expression, but decreased N-cadherin expression when compared with control group. Treatment of lidocaine increased TRAF3 protein expression, but decreased p-p65 protein expression in ES-2 and SKOV3 cells. Conclusion: We demonstrated that lidocaine inhibited cell proliferation, induced apoptosis, and inhibited EMT in ovarian cancer cells via regulating TRAF3/NF-κB pathway.

scholarly journals Effect of Histone Deacetylase HDAC3 on Cytokines IL-18, IL-12 and TNF-α in Patients with Intrahepatic Cholestasis of Pregnancy

2017 ◽  
Vol 42 (4) ◽  
pp. 1294-1302 ◽  
Author(s):  
Yong Shao ◽  
Jing Chen ◽  
Jiao Zheng ◽  
Cai-Ru Liu
Keyword(s):  
Protein Expression ◽  
Histone Deacetylase ◽  
Intrahepatic Cholestasis ◽  
Serum Levels ◽  
Hepatic Function ◽  
Control Group ◽  
Intrahepatic Cholestasis Of Pregnancy ◽  
Tnf Α ◽  
Significant Difference ◽  
Cholestasis Of Pregnancy

Background/aims: The pathogenesis of intrahepatic cholestasis of pregnancy (ICP) is poorly understood. Objective: This study aimed to explore the possible effect of HDAC3 (histone deacetylase) on cytokines IL-18, IL-12 and TNF-α in ICP. Methods: Serum levels of cytokines IL-18, IL-12 and TNF-α, bile acids and hepatic function parameters were measured. The expression of HDAC3 in the placenta was determined by immunohistochemistry (IHC), western blotting and RT-PCR. Results: IL-18, IL-12 and TNF-α serum levels were significantly higher in the severe ICP group than in the mild ICP group and the control group, and the difference between the mild ICP group and control group was not significant. HDAC3 protein expression was identified in the nucleus of the placental trophoblast by IHC. HDAC3 mRNA and protein expression were significantly lower in the ICP groups (mild ICP and severe ICP groups) than in the control groups, and no significant difference was found between the mild ICP and severe ICP groups. Conclusions: The low expression of HDAC3 and overexpession of inflammatory cytokines (IL-18, IL-12 and TNF-α) in ICP may be involved in liver cell apoptosis. We suspect that HDAC3 may play an important role in the pathophysiology of ICP.

scholarly journals PO-082 16-Week high intensity interval training does not alter LKB1 and AMPKα protein in Rats Liver

2018 ◽  
Vol 1 (3) ◽  
Author(s):  
Jia Shao ◽  
Hao Su ◽  
ZhongYe Jiang ◽  
ZhenXing Kong ◽  
GuoHuan Cao ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Intensity ◽  
Interval Training ◽  
Analysis Data ◽  
Training Intervention ◽  
Control Group ◽  
High Intensity Interval Training ◽  
Aging Rats ◽  
Significant Difference ◽  
High Intensity Interval

Objective Liver, as one of the most important organs involved in lipids and glucose metabolism, yet no study has examined the response of liver kinase B1 (LKB1) and AMP-activated protein kinase α(AMPKα) signaling after high intensity interval training. This study aims to evaluate the effect of 16-week high intensity interval training intervention on the expression of LKB1、AMPKα in liver of aging rats. Methods 8 -month-old male Wistar rats（n=40）were randomly divided into control group (C) and HIIT group (H). Group H with 70%-90%-50%VO2max intensity training for 50min/ day, 5 days / week, lasted for 16 weeks. Rats were killed on 0, 8 and 16 weeks. We examined the protein expression of LKB1 and AMPKα in liver. Proteins were analyzed by western blot analysis. Data are mean±SD; for ANOVA, p<0.05 was significant. Results The AMPKα levels in group C and group H increased with time and there was no significant difference between the groups. The content of LKB1 in group C and group H both increased first and then decreased, but there was no significant difference between the groups. Conclusions 16-week high intensity interval training intervention had no effect on LKB1, AMPKα protein expression in aging rats.

scholarly journals Effect of Yiqi Huoxue Granules on Platelet Activation Induced by Thrombin

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Zhen Lei ◽  
Shuibo Gao ◽  
Xinzhou Wang ◽  
Haixia Gao ◽  
Yongjun Han ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Protein Expression ◽  
Intracellular Calcium ◽  
Platelet Activation ◽  
Calcium Concentration ◽  
Control Group ◽  
Aggregation Rate ◽  
Thrombosis Model ◽  
Significant Difference ◽  
Induced Changes

Objective. To study the effects of Yiqi Huoxue (YQHX) granules on platelet activation and aggregation induced by thrombin. Methods. The effect of YQHX on platelet aggregation rate was detected by platelet aggregation instrument; the effect of YQHX on thrombosis time was observed by the mouse mesentery thrombosis model. DAMI cells were induced to transform into platelet-like granules using PMA, and the effects of SCH (PAR-1 inhibitor) on thrombin-induced changes in platelet intracellular calcium concentration, PAR-1 protein expression, and phosphorylation of MAPK were examined. Results. Compared with the control group, the platelet aggregation rate, PAR-1 protein expression, phosphorylation of ERK1/2, and p38 protein in the YQHX group decreased ( P < 0.05 ), and there was no significant difference between the YQHX + SCH group and YQHX group ( P > 0.05 ). Conclusion. YQHX suppresses the platelet activation induced by thrombin by inhibiting PAR-1 expression.

scholarly journals Regulation of NSCLC Cell Proliferation By MARCH7 Via the NF-κB and Wnt/β-Catenin Signaling Pathways

2022 ◽  
Author(s):  
Xiaomi Lu ◽  
Lili Shao ◽  
Ye Qian ◽  
Sixun Zhong ◽  
Jinhong Chen ◽  
...  
Keyword(s):  
Western Blot ◽  
Signaling Pathway ◽  
Reporter Gene ◽  
Nude Mice ◽  
A549 Cells ◽  
Luciferase Reporter ◽  
Clinicopathological Parameters ◽  
G1 Phase ◽  
Gene Assay ◽  
Nsclc Patients

Abstract The aim of the study was to explore the role of the E3 ubiquitin ligase MARCH7 in the development of non-small-cell lung cancer (NSCLC) and to explore the underlying molecular mechanism.Western blot and immunohistochemistry results showed that the expression of MARCH7 in NSCLC cancer tissues was higher than that in paracancerous tissues. Tissue microarray staining results and clinicopathological parameters of NSCLC patients revealed that MARCH7 expression was closely related to TNM stage, degree of tumor differentiation and lymph node metastasis of NSCLC patients. Furthermore, univariate and multivariate analyses and survival curve analysis showed that high expression of MARCH7 was associated with poor prognosis.In vitro, siRNA was constructed and transfected into A549 cells to inhibit the expression of MARCH7. The CCK-8 assay indicated that the growth rate of tumor cells in the interference group was reduced. The number of colonies and cells in the interference group decreased in the plate clone formation experiment. Flow cytometry showed that G0/G1 phase cells were predominantly increased after blocking endogenous MARCH7 expression, and G0/G1 phase arrest occurred in A549 cells. The reporter gene activity of the NF-κB signaling pathway and Wnt/β-catenin signaling pathway was reduced, as validated by a double luciferase reporter gene assay. Western blot analysis showed that the expression of NF-κB P50, NF-κB P65 and β-catenin was decreased, while the expression of E-cadherin was elevated.In vivo, MARCH7-overexpressing virus was constructed and transfected into A549 cells and then subcutaneously injected into nude mice. It was demonstrated that the tumor volume was significantly larger in the MARCH7 overexpression group than in the control nude mice during the same period. Elevated expression of PCNA and Ki-67 was observed in the tumor mass of the MARCH7 overexpression group, as measured by immunohistochemical analysis, accompanied by enhanced levels of NF-κB P50, NF-κB P65 and β-catenin, as detected by Western blot. These results provide a new idea for the experimental basis for the treatment of NSCLC in the future.

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