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@#Objective - - （BCL2L2）- （ ） To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A （PABPN1） （ ） binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the Methods ） ， related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as -， - - low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid （ ）， （ ） ， MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without ， BCL2L2-PABPN1 toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was - （ - ） ） （ ） detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing 基金项目：国家自然科学基金（ ）； 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目 82160607 2021 （ ） 202101AY070001-054 作者简介：施雅（ —），女，在读大学本科生，主要从事劳动卫生与环境卫生学研究；尹锦瑶（ —），女，在读劳动卫生与环境卫 2001 1995 生学硕士研究生，主要从事劳动卫生与环境卫生学研究；施雅和尹锦瑶为共同第一作者 通讯作者：何越峰教授，博士研究生导师，- ： E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 ， ， ， · · 2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523 BCL2L2-PABPN1， - fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group - - BCL2L2-PABPN1 and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After ， BCL2L2-PABPN1 - culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell - survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection ， （ ） ， method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression - Results ） level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of BCL2L2-PABPN1 （P ） BCL2L2- in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of PABPN1 - ， - - in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure （ P ） BCL2L2-PABPN1 ， groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA （ P ） BCL2L2-PABPN1 group and DMA group all <0.05 . The relative expression of showed no significant difference between ， （ P ） ） BCL2L2-PABPN1 control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell - - - （ P ） survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 . ， （P ） However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of - Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and - - ， - siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho ， - - ， - - （ P ） p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 , - - P and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared - Conclusion with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of BCL2L2-PABPN1 fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated BCL2L2-PABPN1 BCL2L2-PABPN1 - metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis BCL2L2 PABPN1 through affecting the synergistic effect of and genes.
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AIM:To investigate the effect of epigallocatechin gallate(EGCG)on the apoptosis of human retinal pigment epithelium(ARPE-19)cells and its mechanism. METHODS:The ARPE-19 cells were cultured in vitro and treated with 0,40,80 and 160 μg/mL EGCG, respectively. At the proposed time of treatment the morphological changes were detected by hoechst 33258 staining. The apoptosis rate was detected by flow cytometry. The expression of apoptosis-related factors B lymphocytoma-2 gene(bcl-2), BCL2-Associated X protein(Bax),caspase-3 and p53 were detected by quantitative RT-PCR and Western blotting.RESULTS: Hoechst 33258 staining showed that the ARPE-19 cells with the increase of EGCG drug concentration, the number of apoptotic cells gradually increased and the apoptotic bodies were observed. Flow cytometry showed that the apoptosis rate increased gradually with the increase of EGCG drug concentration. The apoptosis rates at 40, 80 and 160 μg/mL were 4.95%±0.071%, 11.75%±0.075% and 21.25%±0.919% respectively, which was significantly different compared with the control group(2.8%±1.556%)(P<0.01), presented with a drug concentration-dependent. The results of quantitative PCR and Western blotting showed that EGCG could significantly up-regulate the expression of apoptosis-promoting factors Bax, caspase-3 and the mRNA and protein expression of p53, and down-regulate the apoptosis-inhibiting factor bcl-2, all of these showed concentration-dependent effects.CONCLUSION:EGCG can obviously induce the apoptosis of ARPE-19 cells. The mechanism is related with the inhibition of bcl-2 and increase the expression of Bax, caspase-3 and p53.
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Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P＜0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.
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Background Pterygium is a relatively common eye disease,but its aetiology and pathogenesis remain uncertain.At present,the study on pterygia focuses on understanding its underlying mechanism.Objective This study was to detect the expression of 8-hydroxy-2'-deoxyguaine (8-OHdG),a marker of oxidative damage of DNA,and bcl-2,a gene related with apoptosis,on the pterygium tissue.Methods Thirty pterygium tissue specimens were obtained during the surgery with the primary pterygium 24 cases and recurrent pterygium 6 cases.In addition,20 normal conjunctival specimens from retinal detachment surgery and strabismus surgery were collected.The expressions of 8-OHdG and bcl-2 in pterygium tissue were detected using immunochemistry and compared with the normal conjunctival tissue.The difference in the expressions of 8-OHdG and bcl-2 among different specimens was compared by x2test,and the relationship between 8-OHdG expression and bcl-2 expression was evaluated by Kappa test.Results The positive expressing rate of 8-OHdG in the pterygium tissue was 62.5％ and 83.3％ in the primary and recrudescence pterygium tissue,respectively,but the expression of 8-OHdG was absent in the normal conjunctiva tissue.No significant difference was found in the positive expressing rate of 8-OHdG between primary and recrudescence pterygium tissue(x2 =0.938,P＞0.05).The bcl-2 expressing rate was 90.0％ and 87.5％ in the primary and recrudescence pterygium tissue,respectively.However,that in the normal conjunctival tissue was absent.No significant difference was seen in the bcl-2 expression rate between primary and recrudescence pterygium tissue (x2=0.833,P ＞ 0.05).Of the 27 pterygium tissue with bcl-2 positive expression,8-OHdG showed the positive expression in 20 specimens,and 3 specimens with the bcl-2 negative response were absent reactive to 8-OHdG,showing insignificant difference between them (P＞0.05).The relationship between 8-OHdG expression and bcl-2 expression was concord in a certein extent (Kappa =0.464).Conclusions The upregulation of 8-OHdG in the pterygium tissue indicates that oxidative damage of DNA plays a role in the development of pterygium.Oxidative damage of DNA caused by ultraviolet may be an upriver factor,which induces raising up of expression of bcl-2 and inhibits the apoptosis of normal cells and further proliferation of the conjunctiva tissue,resulting in the genesis and development of pterysium.
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Objective To study the effects of alpinetin on apoptosis of Hep3B cells and explore the related mechanism.Methods Hep3B cells were cultured in vitro,treated with alpinetin; RT-PCR and Western blot was used to detect the mRNA and protein levels of Bcl-2; MTT assay was used to detect the cellular growth inhibitory rate; Annexin V-FITC/PI double staining was used to detect the apoptosis rate of cells; Mitochondrial membrane potential was analyzed by flow cytometry; Western blot was used to detect protein expression of Caspase-3,9 and Cytochrome C ; the experiment was carried out in four groups:control group,high dosage of alpinetin group,middle dosage of alpinetin group and low dosage of alpinetin group.Results The expression of Bcl-2 in Hep3B cells were decreased by alpinetin.After treated with different dosages of alpinetin (40,80,120 μmol/L),the apoptotic inhibitory rate detected by MTT were 6.38％ ± 1.32％,21.58％ ± 1.97％ and 43.18％ ± 3.89％,significantly higher than those in control group (tlowdose =13.01,tmiddle dose =15.12,thighdose =14.79,average P ＜ 0.01) ; the expression of mitochondrial membrane potential green fluorescence protein (GFP) were 18.93％ ± 2.3％,31.11％ ± 2.67％ and46.06％ ± 2.95％,significantly higher than those in control group (tlow dose =16.70,tmiddle dose =31.38,thigh dose =48.15,average P ＜ 0.01).Western blot analysis showed that the expression of Caspase-3,9 andCytochrome C in cytoplasm significantly was higher than those in control group(Caspase-3:llow dose =11.94,tmiddle dose =10.18,thigh dose =18.82,average P ＜0.01; Caspase-9:tlow dose =15.11,tmiddle dose =20.41,thish dose =21.25,average P ＜0.01; Cytochrome C:tlow dose =15.11,tmiddle dose =28.47,thigh dose =16.01,average P ＜ 0.01).while that Cytochrome C in mitochondria significantly lower than those in control group (tlow dose =16.70,tmiddle dose =12.00,thighdose =27.61,average P ＜ 0.01).Conclusions Alpinetin promotes apoptosis of human hepatic cancer cells Hep3B by down-regulating Bcl-2,probably through mitochondrial pathway.
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Objective: To investigate the expressions of B7-H1 and Bcl-2 proteins in human colorectal cancer, and to explore the association of the expressions of B7-H1 and Bcl-2 with clinicopathological features. Methods: The expressions of B7-H1 and Bcl-2 proteins were detected by immunohistochemistry in colorectal cancer tissues and their corresponding para-cancerous normal tissues from 57 cases of colorectal cancer. The association of the expressions of B7-H1 and Bcl-2 with the clinicopathological features was analyzed. Results: The positive expression rates of B7-H1 and Bcl-2 in colorectal cancer tissues were both higher than those in para-cancerous normal tissues (P <0.05). Both of Bcl-2 and B7-H1 expressions in colorectal cancer tissues were negatively correlated with the degree of differentiation of colorectal cancer (P <0.01). The positive expression rate of B7-H1 was significantly elevated in patients with lymph node metastasis (Dukes' stage C-D) compared to the patients without lymph node metastasis (Dukes' stage A-B) (P <0.05). For the patients with colorectal cancer classified as well or moderately differentiated, the expression rate of Bcl-2 in patients with negative expression of B7-H1 was significantly higher than that in patients with positive expression of B7-H1 (P <0.01). Short-term followup showed that the B7-H1 was expressed in colorectal cancer tissues from the patients died of colorectal cancer. Conclusion: The expression of B7-H1 was correlated with the invasion and metastasis of colorectal cancer as well as the survival. The detection of B7-H1 combined with Bcl-2 may have an important clinical significance in the diagnosis, treatment and prognostic prediction for patients with well/moderately differentiated colorectal cancer. Copyright © 2012 by TUMOR.
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@# Objective To observe the effect of porphyra yezoensis polysaccharide (PYP) on cerebrum tissue of diabetic rats and apoptosis. Methods 60 Wistar rats were randomly divided into normal control group, diabetic model group, and treated groups respectively with PYP (0.25 g/kg × d), PYP (0.5 g/kg × d), and PYP (1 g/kg × d). The cerebrum pathologic changes were observed by HE staining method, and the number of apoptotic cells was observed by electrophoresis and the mRNA expression of p53 and bcl-2 was confirmed by RT-PCR. Results PYP improved the cerebrum pathologic changes of diabetic rats. With comparison to the diabetic model group, the cerebrum pathologic change in the treated groups also improved. And the number of apoptotic cells, the expression of p53 in cerebrum tissue decreased and expressions of bcl-2 rose. Conclusion PYP can decrease the expression of p53 in cerebrum tissue and rise expressions of bcl-2 so as to protect brain tissues from cerebral ischemia.
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Objective To investigate the expression and clinical significance of bcl-2 and NF-κB/ p65 in different subgroups of diffuse large B-cell lymphoma (DLBCL). Methods Immunohistochemical staining was used to detect the expressions of CD10, bcl-6, and MUM-1 in the DLBCL patients. According to immunohistochemical algorithm of Hans et al. DLBCL were subdivided into GCB and non-GCB/ABC subgroups and the expressions of bcl-2 and NF-κB/p65 were detected. The expressions of bcl-2 and NF-κB/ p65 in GCB DLBCL were compared with that in ABC DLBCL,and the correlation of bcl-2 and NF-κB/p65 expressions with survival in the two major subgroups of DLBCL were analyzed. Results The expression rates of bcl-2 and NF-κB/p65 proteins in DLBCL were 67.1% and 77.1%, and there was significant correlation between them. The expression rates of bcl-2 and NF-κB/p65 were 52.0 % and 56.0 % in GCB DLBCL, but were 75.6 % and 88.9 % in ABC DLBCL. The expression rates of two proteins were higher in ABC DLBCL than in GCB DLBCL. There was no significant correlation between bcl-2 and NF-κB/p65 protein expressions and overall survival within the GCB DLBCL subgroup, but bcl-2 and NF-κB/p65 expressions had a significant effect on overall survival within the ABC subgroup. Conclusion bcl-2 and NF-κB/p65 expressions are associated with poor survival in the ABC subgroup only. Hence, the significance of bcl-2 and NF-κB/p65 protein expressions should be assessed in the context of DLBCL subgroups in the future.
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Objective To investigate the protein and gene expression of bcl-2, bcl-6 in diffuse large B-cell lymphoma (DLBCL). Methods 73 cases of DLBCL were selected for study using the Envision immunohistochemistry method with a panel of antibodies CD3, CD10, CD20, bcl-6, bcl-2, MUM-1. The bcl-2gene expression in 57 of 73 cases with chromosome translocation t (14; 18), breakage and amplification of 3q27 chromosome in 54 of 73 cases were detected by fluorescence in situ hybridization (FISH) method. Results The percentages of tumor cells expressing CD10, bcl-6, MUM-1, bcl-2 were separately 15.1%, 38.4 %, 71.2 %, 79.2 %. t (14;18) chromosomes were detected in 16 of 57 cases (28.1%). The expression of bcl-2 protein have significantly correlated with immunophenotype subtype (P=0.035), and t (14;18) was significantly correlated with the prognosis (P=0.045). There were no association between the expression of bcl-2 protein and t (14;18)(P=0.710). 11 of 54 cases were presented with 3q27 chromosomal breakage (20.4 %), and 14 cases were chromosomal amplification (25.9 %). The prognosis of cases with positive bcl-6 protein was better than that with negative protein obviously. There was no relationship between bcl-6 and 3q27 chromosomal breakage or amplification (P=1.000). Conclusion The expression of bcl-2, bcl-6 protein and gene were different events and had the different significance on DLBCL. The expression of bcl-2 protein was a prognostic marker correlated with immunophenotype subtype, and GCB type with the positive expression of bcl-2 protein had the poor prognosis. Conversely, t(14;18) was an independent event for the prognosis, and the positive expression have the poor prognosis. Patients who require the target therapy should be detected for the t(14;18). The expression of bcl-6 protein was beneficial to the judgment of DLBCL prognosis, it could be an independent factor of the prognosis. 3q27 chromosomal breakage may be a hint to the poor prognosis.
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Objective To study the expression and its significance of bcl-2 associated death (bad) gene in human optic nerves from traumatic atrophic eyeballs. Methods The optic nerves from 8 normal human donor eyes and 31 traumatic atrophic eyes were studied by immunohistochemistry technique. Results Bad protein was positively expressed in the normal optic nerve myelin sheath and residual myelin portions of optic nerve tissues from traumatic atrophic eyes. The expression of bad protein in the residual portions of myelin sheath was stained significantly stronger than that in normal optic nerves (P0 05). Conclusion Bad might possess the function of promoting the optic nerve atrophy processes in traumatic atrophic eyes.
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[Objective] To explore the genetic mechanism of acupoint catgut embedment in inhibiting hippocampal neuron apoptosis in rats with status epileticus (SE). [Methods] Forty SD rats were randomized into 5 groups: blank control (A), model (B), dilantin (C), routine acupuncture (D) and acupoint catgut embedment (E). Rat models of SE were established by intra-abdominal injection of penicillin. With immunohistochemical method, the expressions of apoptosisrelated genes of c-jun and bcl-2 were observed in the vulnerable neurons of the hippocampus 24 hours after modeling. [Results] Compared with SE model group, the expression of c-jun was decreased and the expression of bcl-2 was increased in group C, D and E 24 hours after modeling; c-jun expression was positively related with the apoptotic index (AI) and bcl-2 expression was negatively related; the effects in group E much differed from those in model group. [Conclusion] The possible mechanism of the acupoint catgut embedment in treating epilepsy is related to the inhibition of the hippocampal neuron apoptosis by prohibiting the expression of c-jun and promoting the expression of bcl-2.
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【Objective】To observe the effect of Sicaotang Decoction which is mainly composed of kidney-nourishing,toxicity-removing and blood-activating herbal medicines on parameters associated with mesenteric proliferation,fibronectin(FN),and Bcl-2 gene expression in lupus nephritis(LN).【Methods】 Sixty LN patients were equally randomized into two groups.The two groups received treatment with hormones and cyclophosphamide,and group Areceived Sicaotang Decoction additionally.The changes of Bcl-2 and Fas expression in CD4+ and CD8+ of T lymphocyte subtypes,serum fibronectin(FN),serum and urine collagen Ⅳ(CⅣ) as well as score of blood stasis were observed.【Results】Sicaotang Decoction reduced the expression of Bcl-2 and Fas expression in CD4+ and CD8+,relieved the proliferation of mesenteric cells,and decreased Bcl-2 positive rate,serum FN level,serum and urine CⅣ levels and score of blood stasis(P
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0.05) was observed. Bcl 2 expression was related with differentiation of GC(P0.05).Conclusion: p53,bcl 2,c erbB 2 may be involved in the development of GC.
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Objective To study the effect of vitamin K3 (VitK3) on the apoptosis of human hepatocellular carcinoma cell line of HEPG2 and its relationship with Caspase-3 and Bcl-2 gene. Method With the treatment of VitK3, time and dosage effect was detected by MTT colorimetry. Appoptosis was observed by morphology, flow cytometry and DNA agarose gelelectrophoresis.Caspase-3 and Bcl-2 mRNA expression was assessed by RT-PCR. ResultVitK3 significantly inhibits HEPG2 cell proliferation and induce apoptosis.Caspase-3 mRNA expression was upregulated. ConclusionVitK3 induces HEPG2 cell apoptosis by way of Caspase-3 gene expression.
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0.05). Conclusions Silencing STAT3 gene can decrease STAT3 gene expressions , increase Bax gene expressions , induce cell apoptosis and suppress the tumor growth in the human breast carcinoma model by the RNAi technology .
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Objective To investigate the relation between the expression of Bcl-2 protein and protection against ischemic neuronal damage by preconditioning with sublethal ischemia .Methods The cerebral ischemia was induced by occlusion of bilateral common carotid arteries .Sixty-three gerbils were divided randomly into four groups:sham operative control group (group A,n=5),ischemic preconditioning control group (group B ;n=6) with a single 2-min cerebral ischemia; ischemia preconditioning group (group PC,n=26) and ischemic control group (group IR,n=26) with 5-min ischemia being induced following 3 days of reperfusion with or without 2-min ischemic preconditioning,then with reperfusion lasting 4 hours (group PC1,n=5;group IR1,n=5),24 hours (group PC2,n=7;group IR2,n=7),72 hours (group PC3,n=7;group IR3,n=7)or 7 days (group PC4,n=7;group IR4 ,n=7) respectively.Paraffin sections of hippocampus were used for Bcl-2 protein immunohistochemical staining.Results In group B,Bcl-2 protein immunoreactivity (the intensity of staining) significantly increased as compared with that in group A(P
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Objective To investigate the influence of ?-interferon (?-IFN) on liver cancer cell line (Hep-G2). Methods Observing the expression of Fas and Bcl-2 by ?-IFN-pretreated Hep-G2 cells via immunohistochemical stain; subsequently treating these cells with adrimysin, and observing the cell death rate and apoptosis of these cell by MTT and electroscopy. Results (1) ?-IFN up-regulating the expression of Fas protein and down-regulating Bcl-2 protein (P