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1.
Article in Chinese | WPRIM | ID: wpr-1017324

ABSTRACT

Objective:To discuss the protective effect of gingerone on the hippocampal neuron HT22 cells after oxygen-glucose deprivation/reoxygenation(OGD/R),and to clarify the related mechanism.Methods:The HT22 cells were cultured,and the OGD/R cell injury model was established by setting the gradient of OGD/R time.The HT22 cells were divided into control group,OGD/R group,OGD/R+ 1 μmol·L-1 gingerone group,OGD/R + 10 μmol·L-1 gingerone group,OGD/R+100 μmol·L-1 gingerone group,and OGD/R+0.2%dimethyl sulfoxide(DMSO)group.The viability of the cells in various groups was detected by CCK-8 assay;the survival rates of the cells in various groups were calculated to determine the optimal drug concentration of gingerone.The cells were divided into control,OGD/R group,OGD/R+ gingerone,and OGD/R+gingerone+nuclear factor erythroid-2-related factor 2(Nrf2)inhibitor(ML385)groups.The cells in OGD/R + gingerone group were treated with gingerone for 4 h before OGD treatment for 8 h followed by reoxygenation for 8 h,and the cells in OGD/R+gingerone+ ML385 group were treated with 10 μmol·L-1 ML385 for 6 h before gingerone treatment.The viability of the cells in various groups was detected by CCK-8 assay;the expression levels of Nrf2,heme oxygenase-1(HO-1),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)proteins in the cells in various groups were detected by Western blotting method;the activity of superoxide dismutase(SOD)and the level of malondialdehyde(MDA)in the cell culture supernatant in various groups were detected by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with control group,the survival rate of the HT22 cells was below 50%after treated with OGD for 8 h and reoxygenation for 8 h,so the HT22 cell OGD/R model was established by treated with OGD for 8 h and reoxygenation for 8 h.Compared with OGD/R group,the survival rates of the cells in OGD/R+different doses of gingerone groups were increased to various extents,and the survival rate of the cells in OGD/R+ 100 μmol·L-1 gingerone group was significantly increased(P<0.01);so 100 μmol·L-1 gingerone was used for the subsequent experiment.Compared with control group,the viability of the cells in OGD/R group was significantly decreased(P<0.01),and the expression levels of Nrf2,HO-1,and Bax proteins in the cells were significantly increased(P<0.01),while the expression level of Bcl-2 protein in the cells was significantly decreased(P<0.05),and the SOD activity in the cell culture supernatant was significantly decreased(P<0.01),and the level of MDA was significantly increased(P<0.01);compared with OGD/R group,the viability of the cells in OGD/R + gingerone group was significantly increased(P<0.01),and the expression levels of Nrf2,HO-1,and Bcl-2 proteins in the cells were significantly increased(P<0.05 or P<0.01),while the expression level of Bax protein in the cells was decreased(P<0.05),the SOD activity in the cell culture supernatant was significantly increased(P<0.01),and the level of MDA was significantly decreased(P<0.01);compared with OGD/R + gingerone group,the viability of the cells in OGD/R + gingerone + ML385 group was significantly decreased(P<0.01),and the expression levels of Nrf2,HO-1,and Bcl-2 proteins were significantly decreased(P<0.01),while the expression level of Bax protein in the cells was significantly increased(P<0.01),the SOD activity in the cell culture supernatant was significantly decreased(P<0.01),and the level of MDA was significantly increased(P<0.05).Conclusion:Gingerone alleviates the oxidative stress damage,and thereby plays an inhibiory effect on the apoptosis of the HT22 neurons by activating the Nrf2/HO-1 signaling pathway after OGD/R.

2.
Tianjin Medical Journal ; (12): 44-50, 2024.
Article in Chinese | WPRIM | ID: wpr-1020968

ABSTRACT

Objective To construct an in vitro"metabolic memory"cell model of HT-22 mouse hippocampal neurons induced by high glucose,and to investigate the effect of"metabolic memory"on apoptosis and histone acetylation in HT-22 cells.Methods HT-22 cells were cultured in high glucose medium(glucose concentration was 55 mmol/L)and conventional glucose medium(glucose concentration was 25 mmol/L),and cells were divided into the control group(NG 4,6 and 8 groups,25 mmol/L glucose was cultured for 4,6 and 8 days,respectively),the high glucose group(HG 4,6 and 8 groups,respectively)and the metabolic memory group(HG2NG2,HG2NG4,HG2NG6,HG4NG2 and HG4NG4 groups,high glucose culture for 2 days to 25 mmol/L glucose culture for 2,4 or 6 days,high glucose culture for 4 days to 25 mmol/L glucose culture for 2 or 4 days).Cell viability was detected by CCK-8 method.The release of lactate dehydrogenase(LDH)in cell culture supernatant was detected,and the optimal time to establish a"metabolic memory"model was selected.Subsequently,cells were divided into the NG4 group,the NG8 group,the HG4 group,the HG4NG4 group and the HG8 group,and the cell morphology of each group was observed by optical microscope.The apoptosis rate was detected by flow cytometry.The activities of deacetylase(HDAC)and histone acetyltransferase(HAT)were detected by enzyme-linked immunosorbent assay(ELISA).Western blot assay was used to detect expression levels of histone deacetylase 4(HDAC4),B lymphocyte tumor 2(Bcl-2),Bcl-2 related X protein(Bax)and Caspase-3 protein.Results The HG4NG4 group was the ideal cell model with high glucose metabolic memory.Cells of the NG4 group and the NG8 group were interwoven into a dense network,growing well,with spindle shaped cells and distinct synaptic structures.However,in the HG4 group and the HG8 group,the cell body became round,synaptic structure disappeared and growth was inhibited.In the HG4NG4 group,the number of cells increased but their morphology was damaged.Results of flow cytometry showed that compared with the NG8 group,the apoptosis rates were significantly increased in the HG8 group and the HG4NG4 group(P<0.05).ELISA results showed that compared with the NG8 group,the expression levels of HDAC4,Bax,and Caspase-3 proteins increased in the HG8 group and the HG4NG4 group,while the expression level of Bcl-2 protein significantly decreased(P<0.05).Compared with the HG8 group,there were no significant differences in protein expression levels of HAT and HDAC in the HG4NG4 group.Western blot reslts showed that compared with the NG8 group,the levels of HDAC4,Bax and Caspase-3 protein increased in the HG8 group and the HG4NG4 group(P<0.05).Compared with the HG8 group,there were no significant differences in protein expression levels in the HG4NG4 group.Conclusion HT-22 mouse hippocampal neurons cultured with 55mmol/L high glucose for 4 days,and then cultured with 25 mmol/L glucose for 4 days are the ideal"metabolic memory"cell model.The mechanism may be related to the increased activity of HDAC,HAT and HDAC4 expression in the hyperglycemic model.

3.
Article in Chinese | WPRIM | ID: wpr-981310

ABSTRACT

This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.


Subject(s)
Male , Animals , Mice , Cisplatin/pharmacology , Carcinoma, Hepatocellular/genetics , MAP Kinase Signaling System , Beclin-1 , Apoptosis , Liver Neoplasms/genetics , Necrosis , Proto-Oncogene Proteins c-bcl-2/metabolism , Cell Line, Tumor , RNA, Messenger/metabolism , Autophagy
4.
Article in Chinese | WPRIM | ID: wpr-993383

ABSTRACT

Objective:To investigate the expression of zinc finger protein 22 (ZNF22) gene in hepatocellular carcinoma (HCC) and its effect on tumor proliferation, apoptosis, invasion and metastasis of HCC.Methods:The expression of ZNF22 in 32 HCC specimens, and 371 HCC samples from the cancer genome atlas database were analyzed. ZNF22 knockdown and negative control SNU-449 and JHH-7 HCC cell lines were constructed. The effects of ZNF22 on HCC cells were observed by cell proliferation assay, plate clone formation assay, apoptosis assay, scratch healing assay, Transwell invasion assay, subcutaneous tumor formation, tail vein injection transfer, and small animal live imaging assay in nude mice.Results:The expression of ZNF22 gene is higher in HCC tissues than in paracellular carcinoma tissues, and the difference was statistically significant ( P<0.001). The growth rate of SNU-449 and JHH-7 cells in ZNF22 knockdown group was lower than that in control group, and the difference was statistically significant ( P<0.001). Compared with negative control group, the clone number formed by SNU-449 cells in ZNF22 knockdown group decreased (26±8 vs. 59±5, P<0.01), the level of apoptosis increased (6.60%±0.22% vs. 2.38%±0.30%, P<0.001), the migration rate decreased (14.47%±6.42% vs. 68.84%±8.01%, P<0.001), and the number of invasive cells decreased (48.00±2.23 vs. 179.00±4.81, P<0.001). There was no obvious tumor growth after subcutaneous injection of JHH-7 cells into nude mice in ZNF22 knockdown group, and the systemic fluorescence expression was lower than that of the negative control group, and the difference was statistically significant ( P<0.05). No metastases were observed on autopsy in knockdown group nude mice. Conclusion:ZNF22 is highly expressed in HCC while knockdowing ZNF22 gene inhibited the growth, proliferation, invasion, metastasis of HCC cells, and induced apoptosis of HCC cells.

5.
Chinese Traditional Patent Medicine ; (12): 3930-3936, 2023.
Article in Chinese | WPRIM | ID: wpr-1028707

ABSTRACT

AIM To explore the effects of rhubarb ethanol extract on autophagy and apoptosis of HT22 cells after hemin-induced injury.METHODS The HT22 cells were divided into the blank group,the model group,and the high,medium and low dose rhubarb ethanol extract groups(25,12.5 and 6.25 μg/mL).After the 24 hours corresponding drug intervention,3 hours induction of 40 μmol/L hemin was used for the modeling of cell cerebral hemorrhage injury.Subsequently,the cells had their cell viability detected by CCK-8 method;their leakage rate of lactate dehydrogenase(LDH)detected by kit;their apoptosis rate detected by flow cytometry;and their apoptosis and autophagy related protein expressions of cysteine protease-3(caspase-3),Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2)and microtubule related protein light chain 3(LC3)detected by Western blot.RESULTS Compared with the normal group,the model group displayed decreased cell viability(P<0.01);increased LDH leakage rate,apoptosis rate and the protein expressions of Bax/Bcl-2,caspase-3,Beclin1 and LC3(P<0.01);and decreased protein expression of P62(P<0.01).Compared with the model group,all of the groups intervened with rhubarb ethanol extract showed increased cell viability(P<0.05,P<0.01),decreased LDH leakage rate,apoptosis rate and protein expressions of Bax/Bcl-2,caspase-3,Beclin1 and LC3(P<0.05,P<0.01),in addition to the decreased protein expression of P62 increased(P<0.05,P<0.01)in the high and middle dose rhubarb ethanol extract groups.CONCLUSION The ethanol extract of rhubarb may protect neurons and reduce cell damage through its efficacy in regulating their autophagy and apoptosis.

6.
Chinese Pharmacological Bulletin ; (12): 1522-1526, 2023.
Article in Chinese | WPRIM | ID: wpr-1013730

ABSTRACT

Aim To study the effect of sodium pyruvate on apoptosis and autophagy of HT22 in mouse hippocampal neuronal cells under hypoxia conditions. Methods HT22 cells were incubated with different concentrations of sodium pyruvate to detect their cellular activity by MTS; iron staining was used to further observe the effect of sodium pyruvate on HT22 cells in mitochondrial metabolism; lysosomal staining was applied to detect the lysosomal changes of sodium pyruvate on HT22 cells; Western blot was used to detect the expression of Bcl-2, Bax and LC3-II/LC3- I proteins. Results To verify whether sodium pyruvate exerted neuroprotective effects on mouse hippocampal HT22 cells through affecting mitochondrial apoptosis and autophagy pathways, which were improved by administration of sodium pyruvate. Conclusions Sodium pyruvate administration under hypoxic conditions can reduce the neuroprotective effect of hypoxic injury by reducing apoptosis and activating autophagy in HT22 cells.

7.
Journal of Experimental Hematology ; (6): 1617-1623, 2023.
Article in Chinese | WPRIM | ID: wpr-1010013

ABSTRACT

OBJECTIVE@#To investigate the effect of miR-22 targeting formin-like protein 2 (FMNL2) on the migration and apoptosis of childhood acute myeloid leukemia (AML) cells.@*METHOD@#Peripheral blood samples from 11 children with AML, 10 children with immune thrombocytopenia, human AML cell lines TF-1a, HL-60, THP-1 and human bone marrow stromal cells HS-5 were used as the research objects. UniCel DxH 800 automatic hematology analyzer detected platelet count, hemoglobin, and white blood cell count in peripheral blood samples, and RT-qPCR detected miR-22 expression in peripheral blood samples and AML cells. HL-60 cells were transfected with LipofectamineTM 2000 kit, the experiments were divided into seven groups: blank (no cells transfected), miR-NC, miR-22 mimics, si-NC, si-FMNL2 , miR-22 mimics+OE-NC and miR-22 mimics+OE-FMNL2 . RT-qPCR was used to detect the expression of miR-22 in each group. Transwell was used to detect cell migration. Flow cytometry was used to detect cell apoptosis. Dual-luciferase reporter gene detection experiments verified the targeting relationship between miR-22 and FMNL2 . Western blot was used to detect the expression of FMNL2 protein.@*RESULTS@#Compared with the control group, the number of leukocytes in the peripheral blood of children with AML was significantly increased (P <0.001), while the concentration of hemoglobin and the number of platelets were significantly decreased P <0.001). The expression level of miR-22 in peripheral blood of children with AML was significantly lower than that in control group (P <0.001). Compared with HS-5 cells, the expression levels of miR-22 in TF-1a, HL-60, and THP-1 cells were significantly decreased (P <0.05), and in HL-60 cells was the lowest. Therefore, HL-60 cells were selected for subsequent experiments. Up-regulation of miR-22 or silencing of FMNL2 could reduce the number of migrating cells and increase apoptosis rate (P <0.05). MiR-22 targeted and negatively regulated the expression of FMNL2 . FMNL2 overexpression reversed the effects of up-regulated miR-22 on migration and apoptosis of HL-60 cells.@*CONCLUSION@#MiR-22 can inhibit the migration and promote apoptosis of HL-60 cells by down regulating the expression of FMNL2 .


Subject(s)
Humans , Child , MicroRNAs/metabolism , Leukemia, Myeloid, Acute/metabolism , Cell Proliferation , Apoptosis , Myeloproliferative Disorders , Cell Movement , Hemoglobins , Cell Line, Tumor , Formins
8.
Article in Chinese | WPRIM | ID: wpr-1038485

ABSTRACT

Objective@#To explore the effects of long non-coding RNA (LncRNA) MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) and its molecular mechanism.@*Methods@#Synovial tissue samples were collected from 37 RA patients and 30 joint trauma patients in our hospital,and the expression levels of MIR22HG and miR-22-5p in synovial tissue were detected by qRT-PCR. RA-FLSs in human was isolated ,cultured and identified in vitro. MIR22HG siRNA interference plasmid (si-MIR22HG) and its negative control plasmid (si-NC) ,miR-22-5p inhibitor and its negative control (inhibitorNC) were transfected into RA-FLSs respectively or simultaneously.The expression levels of MIR22HG and miR- 22-5p were detected by qRT-PCR. CCK-8 was used to detect the proliferation activity of cells in various groups. Annexin Ⅴ- FITC / PI was used to detect the apoptosis rates of cells in various groups.ELISA was used to detect the levels of TNF-α , IL-1 β and IL-6 in the supernatant of cells in various groups.Western blot was used to detect the protein expression levels of Bcl-2,Bax and Cleaved caspase-3 of cells in various groups.The targeting relationship between MIR22HG and miR-22-5p was verified by dual luciferase reporter gene assay. @*Results @#Compared with joint trauma patients,the expression level of MIR22HG in synovial tissues of RA patients increased (P<0. 05) , while the expression level of miR-22-5p decreased (P<0. 05) .Interference with MIR22HG inhibited the proliferation activity of RA-FLSs,decreased the levels of TNF-α , IL-1 β and IL-6 in cell supernatant and the protein expression level of Bcl-2 in cells (P<0. 05) ,and increased the apoptosis rate,the expression level of miR-22-5p and the protein expression levels of Bax and Cleaved casepase-3 (P <0. 05 ) .However,inhibition of miR-22-5p expression reversed the effects of MIR22HG gene silencing on proliferation,apoptosis and inflammation of RA-FLSs (P<0. 05) .Dual luciferase reporting assay showed that miR-22-5p was a potential downstream miRNA target of MIR22HG.@*Conclusion@# MIR22HG is highly expressed in synovial tissues of RA patients,and it may promote the proliferation and the inflammatory response of RA-FLSs and inhibit cell apoptosis by down regulating the expression of miR-22-5p.

9.
Asian Journal of Andrology ; (6): 414-421, 2020.
Article in Chinese | WPRIM | ID: wpr-842452

ABSTRACT

The apoptosis that occurs in the immature testis under physiological conditions is necessary for male germ cell development, whereas improper activation of apoptosis can impair spermatogenesis and cause defects in reproduction. We previously demonstrated that in mice, the makorin-2 (Mkrn 2) gene is expressed exclusively in the testis and its deletion leads to male infertility. To understand the potential molecular mechanism, in this study, we found that levels of apoptosis in the testis were abnormally high in the absence of Mkrn 2. To identify specific gene(s) involved, we performed digital gene expression profiling (DGE) and pathway analysis via gene set enrichment analysis (GSEA) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and we found that MKRN2 inhibits p53 apoptosis effector related to PMP22 (PERP) expression and that levels of the protein in sperm samples have an inverse correlation with infertility levels. GSEA additionally indicated that PERP is a negative regulator of spermatogenesis and that its ectopic expression induces male infertility. Further, Gene Expression Omnibus (GEO) dataset analysis showed that p53, upstream of PERP, was upregulated in oligoasthenoteratozoospermia (OAT). These observations suggest that Mkrn 2 is crucial for protecting germ cells from excessive apoptosis and implicate Mkrn 2-based suppression of the p53/PERP signaling pathway in spermatogenesis and male fertility.

10.
Chinese Pharmacological Bulletin ; (12): 519-524, 2019.
Article in Chinese | WPRIM | ID: wpr-857368

ABSTRACT

Aim: To investigate the effect of astragaloside IV on apoptosis of HT22 after oxygen and glucose deprivation/reoxygenation by regulating autophagy. Methods: HT22 cells were randomly divided into control, model, DMSO, AS-IV, AS-IV + N, AS-IV + 3-MA, 3-MA and Rapa group. Except for control group, cells in other groups were reoxygenated after 6 h of oxygen and glucose deprivation. Invertded microscope was employed to observe cell morphology. CCK-8 method was used to test cell survival rate. LDH method was applied to detect cell damage, and Bax, Bcl-2 immunofluorescence was used to detect apoptosis. Results: Compared with control, the synapses of cell bodies decreased, the cells shrank, the number of intercellular connections decreased, cell viability was significantly reduced, and LDH leakage and Bax/Bcl-2 significantly increased in model group (P <0. 01). Compared with model group, cell viability in AS-IV group and Rapa group markedly increased, and LDH leakage and Bax/ Bcl-2 significantly decreased (P < 0. 01); cell viability was significantly down-regulated, and Bax/Bcl-2 was markedly up-regulated in 3-MA group (P < 0. 01). Astragaloside IV +3-MA group had no significant difference compared with model group. Conclusion: Astragaloside IV protects OGD/R HT22 by inhibiting apoptosis via activating autophagy.

11.
Article in Chinese | WPRIM | ID: wpr-697576

ABSTRACT

Objective To investigate the expression of USP22 in cervical cancer cells,and the effects of USP22 on cell proliferation and chemosensitivity to cisplatin in cervical cancer cells. Methods Quantitative real-time polymerase chain reaction(qRT-PCR)was conducted to detect the expression of USP22 mRNA in cervical cancer cells.Cell count kit-8,flow cytometry,and tumorigenesis were performed to detect the effects of USP22 on the proliferation,apoptosis,cycle cycle,and chemosensitivity of cervical cancer cells to cisplatin in vitro and in vivo. Results USP22 was upregulated in cervical cancer cells compared to human immortalized epidermal cells. Cell proliferation was inhibited,cell apoptosis was promoted,cell cycle was arrested in G1 stage and chemosensi-tivity to cisplatin was enhanced in cervical cancer cells transfected with USP22-siRNAs.Furthermore,tumorigene-sis assays proved tumor growth was inhibited and chemosensitivity to cisplatin was enhanced when USP22 was silenced in HeLa cells. Conclusion USP22 expression is upregulated in cervical cancer cells,and USP22 could promote cell proliferation and inhibit chemosensitivity to cisplatin in cervical cancer cells.

12.
Chinese Pharmacological Bulletin ; (12): 1600-1605, 2017.
Article in Chinese | WPRIM | ID: wpr-667307

ABSTRACT

Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ~ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08%.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.

13.
Chinese Pharmaceutical Journal ; (24): 280-283, 2016.
Article in Chinese | WPRIM | ID: wpr-859204

ABSTRACT

OBJECTIVE: To investigate the effects of Rhaponticum uniflorum extract (RUE) on angiogenesis and apoptosis of H22 transplanted tumor tissue in mice. METHODS: Mice bearing H22 hepatoma cells were randomly assigned into 5 groups: model, high, medium and low dose RUE, and 5-fluorouracil (5-Fu) group. The intervention was lasted for 10 d. The pathological changes were detected with hematoxylin & eosin (HE) staining, the apoptosis of hepatoma cells were determined by DNA laddering method, the microvessel densities (MVD) were detected using immunohistochemical assay, and the protein expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2) and hypoxia-inducible factor-1α (HIF-1α) were detected by Western blot method. RESULTS: RUE treatment reduced the cell proliferation, aggravated the necrosis of transplanted tumor tissue, reduced DNA fragmentation of H22 hepatoma cells, decreased MVD of tumor tissue, and down-regulated the protein expression of VEGF, VEGFR2 and HIF-1α of the transplanted tumor tissue, as compared with the model group. CONCLUSION: RUE could exhibit anti-angiogenic and pro-apoptotic effects against H22 hepatoma cells in mice, and its anti-angiogenic mechanism is probably related to down-regulation of VEGF, VEGFR2 and HIF-1α proteins.

14.
China Oncology ; (12): 29-34, 2014.
Article in Chinese | WPRIM | ID: wpr-439561

ABSTRACT

Background and purpose:Quinonoids can change the cell cycle distribution of tumor cells, and effect the radiosensitizing. This study aimed to investigate the radiosensitization effects, cell cycle and apoptosis of cryptotanshinone on H22 hepatoma-bearing mice. Methods:The mouse hepatoma H22 model was established, then divided into blank control group, irradiation alone group, high dose of cryptotanshinone group, cryptotanshinone (low, medium and high)+IR groups. After irradiated, observed the growth of tumor’s conditions, record epigenetic tumor irradiation time, calculated the delay time of tumor growth and enhancement factor (EF). After 22 days, mice were killed, stripped tumor, and calculated the inhibition rate. The cell cycle distribution and apoptosis were measured by lfow cytometry. Results:Cryptotanshinone (low, medium and high) groups inhibited the tumor growth better than the blank group, and had the signiifcant radiosensitizing effect. The enhancement factor was 1.22, 1.43,2.19, respectively. Cells were treated with cryptotanshinone which had significant effects on cell cycle, and induced apoptosis, which indicated signiifcant G2/M phase arrest and a decrease in S phase. Conclusion:Cryptotanshinone inhibited the tumor growth and had the radiosensitizing effects on H22 hepatoma-bearing mice. One of the mechanism may be that it might make significant G2/M phase arrest and S phase decreased, and induced apoptosis. So cells were more sensitive to radiation.

15.
Journal of Leukemia & Lymphoma ; (12): 706-711, 2013.
Article in Chinese | WPRIM | ID: wpr-467670

ABSTRACT

Objective To investigate the effects of overexpression of heat shock protein 22(HSP22) in hematopoietic malignant tumor cell lines.Methods A lentiviral system was used to mediate transduction of HSP22 complementary DNA-containing expression vector or empty vector into K562 and Namalwa cells.The transduction effeciency was tested by fluorescence microscope scan and flow cytometry.Semi-quantitative RT-PCR and Western blot were used to identify the expression levels of HSP22 mRNA and protein.Growth curve analysis,cell cycle analysis,colony-forming assay,tumor growth in nude mice and apoptosis analysis were used to evaluate the role of HSP22 in K562 and Namalwa cells.Results Lentivector expression systemmediated delivery of HSP22 into K562 and Namalwa cells can inhibit colony forming of K562 and Namalwa cells,the average numbers of colonies per well were 108,72,125 and 80 for K562-V,K562-H,Namalwa-V and Namalwa-H respectively (P =0.000 16 and 0.000 37 for K562 and Namalwa respectively).HSP22 transduction can also inhibit proliferation of Namalwa cells in vitro (P =0.015,0.042 and 0.048 for day 5,6 and 7 respectively) and K562 cells in vivo (P =0.022 for day 21).No significant difference in cell cycle and apoptosis was found in K562 and Namalwa cells compared with controls (all P > 0.05).Conclusion Overexpression of HSP22 could inhibit the growth of hematopoietic malignant tumor cell lines K562 and Namalwa.

16.
Article in English | WPRIM | ID: wpr-812327

ABSTRACT

AIMS@#To investigate the antitumor effects of extracts from Oxytropis falcata on human hepatocellular carcinoma SMMC-7721 cells in vitro and in transplanted murine H22 tumors in vivo.@*METHODS@#Cell proliferation, cell cycle distribution and apoptosis in SMMC-7721 cells were determined and tumor growth inhibition in H22 tumors was investigated. Cell cycle distribution was analyzed by flow cytometry with propidium iodide (PI) and Annexin V-FITC/ PI double staining.@*RESULTS@#MTT assay revealed that essential oil and flavonoids of O. falcata (named EOOF and FOF) inhibited proliferation of SMMC-7721 cells in a dose-dependent manner. The IC50 value of EOOF and FOF were 0.115 and 0.097 mg·mL(-1), respectively. Cell cycle was arrested at G(1) phase, and induction of apoptosis occurred in SMMC-7721 cells when subjected to FOF. Growth inhibition in H22 solid tumors transplanted mice was significantly pronounced after being treated with FOF, and the inhibition ratio were 56.1% and 70.8% at the concentration of 30 and 60 mg·kg(-1).@*CONCLUSION@#The results suggest that FOF promotes apoptosis in SMMC-7721 cells and inhibits H22 tumor growth, resulting in a potential antitumor effect on hepatic cancer.


Subject(s)
Animals , Humans , Male , Mice , Antineoplastic Agents, Phytogenic , Apoptosis , Carcinoma, Hepatocellular , Drug Therapy , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Growth Inhibitors , Liver Neoplasms , Drug Therapy , Mice, Inbred ICR , Oxytropis , Chemistry
17.
Article in Chinese | WPRIM | ID: wpr-855066

ABSTRACT

Objective: To investigate the antitumor effect of volatile oil from Sinapis Albae Semen (VOSAS) on H22-bearing mice and to determine the mechanism. Methods: To establish the H22 implanted hepatocellular carcinoma animal model which was used to analyze the effect of VOSAS on the growth of transplanted tumor. Mice were divided into five groups 24 h after modeling: model, cytoxan (CTX, 25 mg/kg) positive control, low-, mid-, and high-dose (20, 40, and 80 mg/kg) VOSAS groups. The mice were ip administered once daily for 10 d. Morphological changes in H22 solid tumor cells were observed by both Hematoxylin-eosin (HE) and acridine orange (AO) staining. The expression of Bax and Bcl-2 in the tumor tissue was determined using immunohistochemistry. Results: VOSAS could inhibit the tumor growth and extend the life span of H22-bearing mice (P < 0.01); and it could also raise the expression of Bax while suppress the expression of Bcl-2; the antitumor effect of VOSAS on H22-bearing mice demonstrated a good dose-effect relationship, but the high-dose group of the volatile oil has obvious toxicity and side effects on the mice. Conclusion: VOSAS could inhibit the growth of H22 tumor cells and the mechanism may be related to up-regulating the expression of Bax and down-regulating the expression of Bcl-2, and the induction of apoptosis.

18.
International Journal of Surgery ; (12): 849-852, 2009.
Article in Chinese | WPRIM | ID: wpr-392063

ABSTRACT

Interleukin-22 (IL-22) is a new kind of eytokine discovered in 2000. The major sources of IL-22 are activated T1 -cells and NK-cells. Tissue cells at outer body barriers, i.e. of the skin, kidney, the di-gestive and respiratory systems all highly express IL-22R or respond to IL-22. IL-22 functions by promoting the anti-microbial defense, inducing phase reactants, protecting against damage and enhancing natural immu-nity. Furthermore, IL-22 mediates the proliferation, differentiation and apoptesis in cancer cells, that gives us a new idea about tumor therapy.

19.
Tumor ; (12): 602-606, 2007.
Article in Chinese | WPRIM | ID: wpr-849522

ABSTRACT

Objective: To investigate the apoptosis-inducing effect of matrine on murine hepatocarcinoma cell line H22 in vivo and in vitro and explore the underlying mechanisms. Methods: The H22 cell apoptosis induced by matrine at the early stage was detected with Annexin V-FITC/PI double staining assay, The expressions of Bcl-2 and Bax proteins in H22 cells as well as in the BALB/c H22 xenograft tumor tissues were detected using immunohistochemical method. Transmission electron microscopy (TEM) was used to observe the ultramicro-structure alterations of H22 xenograft tumor cells in BALB/c mice. The effect of matrine on the kinetics of tumor growth after subcutaneous injection of H22 cells in BALB/c mice was observed and the tumor inhibition rate was calculated. Results: Annexin V staining detected early apoptosis of H22 cells after treatment with matrine 1.0 mg/mL and 1.5 mg/mL for 48 h. The apoptotic rates were 11.71% and 17.86%, respectively, both of which higher than that of control groups (P <0.05). The tumor inhibition rate was above 60% after matrine treatment. Immunohistochemistry analysis revealed that matrine increased Bax protein expression and reduced Bcl-2 protein expression in both H22 cells in vitro and xenograft tumor tissues in vivo. TEM demonstrated the existence of apoptotic cells and apoptotic bodies in H22 xenograft tumor tissues after matrine treatment. Conclusion: Matrine significantly suppresses tumor growth and induced apoptosis both in vitro and in vivo. The apoptosis-inducing effects is related with up-regulation of Bax protein and down-regulation of Bcl-2 protein.

20.
Article in Chinese | WPRIM | ID: wpr-566270

ABSTRACT

[Objective]To study the inhibitory effect of the extract of Rhizoma Amorphophallus,we set up the Kunming mices which were vaccinated with H22 tumor as experimental model,and explore the possibility mechanism of them.[Method]We adopted the mices vaccinated with H22 tumor as experimental model to observe the influence of different extracts on the antitumor ratio,and the cooperating antitumor effect together with 5-Fu.Using flow cytometry to detect the effect of petroleum ether extract on apoptotic cells.Using immunohistochemical method to observe the influence of this extract to the expression of Survivin and Bax(Bcl-2 associated protein x).[Result]The extract of petroleum ether of Rhizoma Amorphophalli had inhibitory effect of the growth on mice vaccinated with H22 tumor,and the possible mechanism was the downregulation of Survivin expression and upregulation of Bax expression to induce apoptosis of tumor cell.

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