Expression of second mitochondria-derived activator of caspases, X-linked inhibitor of apoptosis protein, and caspase-3 in pituitary adenomas

Studies concerning correlations between pituitary adenomas and cell apoptosis have mainly focused on upstream apoptosis signaling, but seldom on downstream mediators. In the present study, second mitochondria-derived activator of caspases （Smac）, X-linked inhibitor of apoptosis protein （XIAP）, and caspase-3 protein were qualitatively analyzed using immunohistochemistry, and quantified by western blot. Smac, XIAP, and caspase-3 mRNA expressions were detected by reverse transcription-PCR. Results showed that XIAP protein and mRNA expressions were greater in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. However, Smac and caspase-3 protein and mRNA expressions were lower in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. In the invasive pituitary adenomas, Smac expression was positively correlated with caspase-3 protein and mRNA expression （Protein： r = 0.55, P 0.01; mRNA： r = 0.50, P 0.01）. Smac and caspase-3 expressions were negatively correlated with XIAP protein and mRNA expression （Protein： r = -0.56, -0.64, P 0.01; mRNA： r = -0.69, -0.67, P 0.01）. However, no significant differences in correlation among Smac, XIAP, and caspase-3 were detectable in noninvasive pituitary adenomas. These data indicated that high expression of XIAP and low expression of Smac and caspase-3 suppressed cell apoptosis and led to enhanced invasiveness of pituitary adenomas. Thus, Smac, XIAP, and caspase-3 may be useful markers in determining the invasive behavior of pituitary adenomas.

Expression of X-linked Inhibitor of Apoptosis Protein and Its Effect on Chemotherapeutic Sensitivity of Bladder Carcinoma

The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemothera- peutic sensitivity of T24 cells.

Targeting X-linked inhibitor of apoptosis protein inhibits pancreatic cancer cell growth through p-Akt depletion

AIM: To determine whether lentivirus-mediated shRNA targeting the X-linked inhibitor of apoptosis protein (XIAP) gene could be exploited in the treatment of pancreatic cancer. METHODS: Human pancreatic cancer cells Panc-1, Mia-paca2, Bxpc-3 and SW1990, infected with lentivirus, were analyzed by real-time polymerase chain reaction (PCR). Western blotting was used to examine XIAP protein levels, survivin and p-Akt to confirm the result of real-time PCR and determine the possible mechanism. The 3-(4,5-cimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) assay was used to measure IC50 to determine chemosensitivity to the chemotherapeutic drugs 5-fluorouracil (5-FU) and gemcitabine. A colony assay, MTT assay and a tumorigenicity experiment were used to study cell proliferation in vitro and in vivo . Caspase-3/7 activity, 4',6-diamidino-2-phenylindole-staining and flow cytometric measurements were used to study apoptosis in SW1990 cells. RESULTS: XIAP proteins were found to be differen- tially expressed among pancreatic cancer cell lines Panc-1, Mia-paca2, Bxpc-3 and SW1990. Data of real-time PCR and Western blotting showed that XIAP was reduced persistently and markedly by lentivirus-mediated shRNA. Downregulation of XIAP by transfection with XIAP shRNA resulted in decreased p-Akt expression. XIAP shRNA also inhibited the growth of pancreatic cancer cells in vitro and in vivo , enhanced drug-induced apoptosis and increased chemosensitivity to 5-FU and gemcitabine. Results also suggest that inhibition of XIAP and subsequent p-Akt depletion may have an anti-tumor effect through attenuating the ability of cancer cells to survive. CONCLUSION: Lentivirus-mediated gene therapy is an attractive strategy in the treatment of pancreatic cancer and justifies the use of lentivirus in pancreatic cancer gene therapy studies.

SMAC exhibits anti-tumor effects in ECA109 cells by regulating expression of inhibitor of apoptosis protein family

BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.

Human antigen R mediated post-transcriptional regulation of inhibitors of apoptosis proteins in pancreatic cancer

AIM To determine the association of human antigen R(HuR) and inhibitors of apoptosis proteins(IAP1, IAP2) and prognosis in pancreatic cancer.METHODS Protein and mRNA expression levels of IAP1, IAP2 and HuR in pancreatic ductal adenocarcinoma(PDAC) were compared with normal pancreatic tissue. The correlations among IAP1/IAP2 and HuR as well as their respective correlations with clinicopathological parameters were analyzed. The Kaplan-Meier method and log-rank tests were used for survival analysis. Immunoprecipitation assay was performed to demonstrate HuR binding to IAP1, IAP2 mRNA. PANC1 cells were transfected with either anti-HuR siRNA or control siRNA for 72 h and quantitative reverse transcription polymerase chain reaction(RT-PCR), western blot analysis was carried out.RESULTS RT-PCR analysis revealed that HuR, IAP1, IAP2 mRNA expression were accordingly 3.3-fold, 5.5-fold and 8.4 higher in the PDAC when compared to normal pancreas(P < 0.05). Expression of IAP1 was positively strongly correlated with HuR expression(P < 0.05, r = 0.783). Western blot analysis confirmed RTPCR results. High IAP1 expression, tumor resection status, T stage, lymph-node metastases, tumor differentiation grade, perineural and lymphatic invasion were identified as significant factors for shorter survival in PDAC patients(P < 0.05).Immunohistological analysis showed that HuR was mainly expressed in the ductal cancer cell's nucleus and less so in cytoplasm. RNA immunoprecipitation analysis confirmed IAP1 and IAP2 post-transcriptional regulation by HuR protein. Following siHuR transfection, IAP1 mRNA and protein levels were decreased, however IAP2 expression levels were increased.CONCLUSION HuR mediated overexpression of IAP1 significantly correlates with poor outcomes and early progression of pancreatic cancer. Further studies are needed to assess the underlying mechanisms.

Baculoviral inhibitor of apoptosis protein repeatcontaining protein 3 delays early Wallerian degeneration after sciatic nerve injury

Wallerian degeneration is a complex biological process that occurs after nerve injury,and involves nerve degeneration and regeneration.Schwann cells play a crucial role in the cellular and molecular events of Wallerian degeneration of the peripheral nervous system.However,Wallerian degeneration regulating nerve injury and repair remains largely unknown,especially the early response.We have previously reported some key regulators of Wallerian degeneration after sciatic nerve injury.Baculoviral inhibitor of apoptosis protein repeat-containing protein 3(BIRC3)is an important factor that regulates apoptosis-inhibiting protein.In this study,we established rat models of right sciatic nerve injury.In vitro Schwann cell models were also established and subjected to gene transfection to inhibit and overexpress BIRC3.The data indicated that BIRC3 expression was significantly up-regulated after sciatic nerve injury.Both BIRC3 upregulation and downregulation affected the migration,proliferation and apoptosis of Schwan cells and affected the expression of related factors through activating c-fos and ERK signal pathway.Inhibition of BIRC3 delayed early Wallerian degeneration through inhibiting the apoptosis of Schwann cells after sciatic nerve injury.These findings suggest that BIRC3 plays an important role in peripheral nerve injury repair and regeneration.The study was approved by the Institutional Animal Care and Use Committee of Nantong University,China(approval No.2019-nsfc004)on March 1,2019.

MG132 Induced Apoptosis Pathway in HL-60 Cells and Impact of Allogeneic Mixed Lymphocyte Reaction

Objective： To investigate the proteasome inhibitor MG132-induced apoptosis pathway in HL-60 cells and the role of allogeneic mixed lymphocyte reaction. Methods： Cell apoptosis was analyzed by flow cytometry. The expressions of p21 protein, p27 protein and p53 protein in HL-60 ceils treated with MG132 were measured by Western blot. The proliferation of, peripheral blood mononuclear cells （PBMNCs） after treatment with 75 Gy irradiated HL-60 cells treated with MG132 was measured with CCK-8. Results： High-dose MG132 induced apoptosis in HL-60 cells. No significant change was observed in MG132-induced apoptosis after inhibiting caspase-8 and caspase-9 pathway. The expressions of p21 protein and p27 protein increased in MG132-induced apoptosis. HL-60 cells treated with low-dose MG132 improved the proliferation of PBMNCs from healthy volunteers. Conclusion： High-dose MG132 induced apoptosis and directly killed HL-60 ceils. MG132 induced apoptosis in a caspase-8- and caspase-9-independent pathway, p21 protein and p27 protein were involved in MG132-induced apoptosis in HL-60 cells. HL-60 cells treated with Low-dose MG132 improved the effect of promoting the proliferation of PBMNCs from healthy volunteers.

Effect of Nimesulide on proliferation and apoptosis of human hepatoma SMMC-7721 cells

AIM: Cyclooxygenase-2 (COX-2) has been suggested to beassociated with carcinogenesis. We sought to investigatethe effect of the selective COX-2 inhibitor, Nimesulide onproliferation and apoptosis of SMMC-7721 human hepatomacells.METHODS: This study was carried out on the culture ofhepatic carcinoma SMMC-7721 cell line. Variousconcentrations of Nimesulide (0、200 μmol/L、300 μmol/L、400μmol/L) were added and incubated. Cell proliferation wasdetected with MTT colorimetric assay, cell apoptosis byelectron microscopy, flow cytometry and TUNEL.RESULTS: Nimesulide could significantly inhibit SMMC-7721cells proliferation dose-dependent and in a dependentmanner compared with that of the control group. Theduration lowest inhibition rate produced by Nimesulide inSMMC-7721 cells was 19.06 %, the highest inhibition ratewas 58.49 %. After incubation with Nimesulide for 72 h, themost highest apoptosis rate and apoptosis index of SMMC-7721 cells comparing with those of the control were 21. 20 %+1.62% vs2.24% +0.26% and21.23+ 1.78 vs2.01+0.23(p＜0.05).CONCLUSION: The selective COX-2 inhibitor, Nimesulide caninhibit the proliferation of SMMC-7721 cells and increaseapoptssis rate and apoptosis index of SMMC-7721 cells. Theapoptoois rate and the apoptosis index are dose-dependent.Under electron microscope SMMC-7721 cells incubated with 300μmol and 400 μmol Nimesulide show apoptotic characteristics.With the clarification of the mechanism of selective COX-2inhibitors, These COX-2 selective inhibitors can become thechoice of prevention and treatment of cancers.

XAF1 mediates apoptosis through an extracellular signal-regulated kinase pathway in colon cancer

Background:XIAP-associated factor 1(XAF1)negatively regulates the function of the X-linked inhibitor of apoptosis protein(XIAP),a member of the IAP family that exerts antiapoptotic effects.The extracellular signal-regulated kinase(ERK)pathway is thought to increase cell proliferation and to protect cells from apoptosis.The aim of the study was to investigate the correlation between the ERK1/2 signaling pathway and XAF1 in colon cancer.Methods:Four human colon cancer cell lines,HCT1116 and Lovo(wildtype p53),DLD1 and SW1116(mutant p53),were used.Lovo stable transfectants with XAF1 sense and antisense were established.The effects of dominant-negative MEK1(DN-MEK1)and MEK-specific inhibitor U0126 on the ERK signaling pathway and expression of XAF1 and XIAP proteins were determined.The transcription activity of core XAF1 promoter was assessed by dual luciferase reporter assay.Cell proliferation was measured by MTT assay.Apoptosis was determined by Hoechst 33258 staining.Results:U0126 increased the expression of XAF1 in a time-and dose-dependent manner.A similar result was obtained in cells transfected with DN-MEK1 treatment.Conversely,the expression of XIAP was down-regulated.Activity of the putative promoter of the XAF1 gene was significantly increased by U0126 treatment and DN-MEK1 transient transfection.rhEGF-stimulated phosphorylation of ERK appeared to have little or no effect on XAF1 expression.Overexpression of XAF1 was more sensitive to U0126-induced apoptosis,whereas down-regulation of XAF1 by antisense reversed U0126-induced inhibition of cell proliferation.Conclusions:XAF1 expression was up-regulated by inhibition of the ERK1/2 pathway through transcriptional regulation,which required de novo protein synthesis.The results suggest that XAF1 mediates apoptosis induced by the ERK1/2 pathway in colon cancer.

JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

瞄准：在 JTE-522-induced apoptosis 调查 mitochondrial 小径的角色并且调查在细胞色素 C 版本， caspase 活动和 mitochondrial 膜潜力(Deltapsim ) 的损失之间的关系。方法：房间文化，数的房间， ELISA 试金， TUNEL，流动 cytometry，西方的污点和 fluorometric 试金被采用在 AGS 房间和相关分子的机制在房间增长和 apoptosis 上调查 JTE-522 的效果。结果：JTE-522 禁止了 AGS 细胞的生长并且导致了 apoptosis。Caspases 8 并且 9 在从 procaspase 和 caspase 活动由劈开产品的外观判定了劈开特定的 fluorogenic 底层的 apoptosis 期间被激活。阐明是否 caspases 的激活 8 和 9 为 apoptosis 正式就职被要求，我们在 apoptosis 上检验了 caspase 特定的禁止者的效果。结果证明 caspase 禁止者显著地禁止了 JTE-522 导致的 apoptosis。另外，与 Rhodamin 123 的举起的减少伴随的细胞色素 C 的 Bax 和 cytosolic 版本的膜 translocation，在 apoptosis 的一个早阶段被检测。而且， Bax translocation，细胞色素 C 版本，和 caspase 9 激活被 Z-VAD.fmk 和 Z-IETD-CHO 堵住。结论：现在的数据显示在 caspases 的激活之间的一个关键协会 8， 9，细胞色素 C 版本， Bax 的膜 translocation， Deltapsim 的损失和在 AGS 房间的 JTE-522-induced apoptosis。

The involvement of p38 MAPK in transforming growth factor β1-induced apoptosis in murine hepatocytes

We reported in this manuscript that TGF-β1 induces apoptosis in AML12 murine hepatocytes, which is associated with the activation of p38 MAPK signaling pathway. SB202190, a specific inhibitor of p38 MAPK, strongly inhibited the TGF-β1-induced apoptosis and PAI-1 promoter activity. Treatment of cells with TGF-β1 activates p38. Furthermore, over-expression of dominant negative mutant p38 also reduced the TGF-β1-induced apoptosis. The data indicate that the activation of p38 is involved in TGF-β1-mediated gene expression and apoptosis.

Isobaric tags for relative and absolute quantitation-based quantitative proteomic analysis of X-linked inhibitor of apoptosis and H2AX in etoposide-induced renal cell carcinoma apoptosis

Background:X-linked inhibitor of apoptosis(XIAP)is a vital factor in the anti-apoptosis mechanism of tumors and is highly expressed in renal cell carcinoma(RCC).However,the mechanism through which XIAP regulates DNA damage repair is unknown.This study investigated the regulatory mechanism of XIAP in etoposide-induced apoptosis in two Caki-1 cell lines with high or low XIAP expression.Methods:The two cell lines were established using RNA interference technology.The differentially expressed proteins in the two cell lines were globally analyzed through an isobaric tags for relative and absolute quantitation-based quantitative proteomics approach.Proteomic analysis revealed 255,375,362,and 5 differentially expressed proteins after 0,0.5,3,and 12 h of drug stimulation,respectively,between the two cell lines.The identified differentially expressed proteins were involved in numerous biological processes.In addition,the expression of histone proteins(H1.4,H2AX,H3.1,H3.2,and H3.3)was drastically altered,and the effects of XIAP silencing were accompanied by the marked downregulation of H2AX.Protein-protein interactions were assessed and confirmed through immunofluorescence and Western blot analyses.Results:The results suggested that XIAP may act as a vital cell signal regulator that regulates the expression of DNA repair-related proteins,such as H2AX,and influences the DNA repair process.Conclusions:Given these functions,XIAP may be the decisive factor in determining the sensitivity of RCC cell apoptosis induction in response to chemotherapeutic agents.

微小RNA-103a-3p通过肿瘤蛋白53调控凋亡抑制剂1/P53对骨质疏松症的影响

目的探讨微小RNA(miR)-103a-3p调控细胞肿瘤蛋白53调控凋亡抑制剂1(TRIAP1)对成骨细胞分化以及去卵巢小鼠骨量的影响。方法MC3T3-E1细胞分为正常对照(NC)组、miR-103a-3p-NC组、miR-103a-3p模拟(mimc)组、miR-103a-3p mimic+TRIAP1-NC组、miR-103a-3p mimic+TRIAP1 mimic组。Real-time PCR检测细胞miR-103a-3p、TRIAP1、P53的mRNA表达,MTT法和流式细胞术检测细胞增殖及凋亡,免疫荧光染色和茜红素染色检测细胞骨架F-actin表达和矿化情况,ELISA检测细胞碱性磷酸酶(ALP)活性。24只雌性小鼠设为sham组、骨质疏松症(OP)组、miR-103a-3p antagonist-NC组和miR-103a-3p antagonist组,每组6只摘取双侧卵巢制备OP模型,sham组仅分离卵巢组织周围脂肪。测定骨组织miR-103a-3p、TRIAP1、P53、ALP、骨钙素(OCN)、骨桥蛋白(OPN)的mRNA表达,microCT测定骨密度(BMD)、骨矿物质含量(BMC),HE染色观察骨组织病理改变。结果细胞转染miR-103a-3p mimic后,miR-103a-3p及P53表达升高、TRIAP1表达降低,细胞增殖降低、凋亡增加,F-actin表达减弱,钙结节数量减少,ALP活性降低(P<0.01);而在增加转染TRIAP1 mimic后,以上miR-103a-3p mimics导致的结果均得到显著逆转(P<0.01)。OP组小鼠骨组织miR-103a-3p、P53表达升高,TRIAP1、ALP、OCN、OPN基因表达降低,BMD、BMC降低,骨组织结构破坏(P<0.05);miR-103a-3p antagonist组小鼠骨组织miR-103a-3p及P53表达降低,TRIAP1、ALP、OCN、OPN基因表达升高,BMD、BMC升高,骨组织结构改善(P<0.05)。结论MiR-103a-3p可介导TRIAP1/P53抑制成骨细胞增殖及矿化,而miR-103a-3p拮抗治疗可减少OP小鼠骨量丢失。

组蛋白去乙酰化酶抑制剂SAHA对慢性间歇性低氧小鼠肝损伤的影响及其机制

目的 探索组蛋白去乙酰化酶抑制剂SAHA对凋亡蛋白Caspase-9/Caspase-3的表达和慢性间歇性低氧小鼠肝损伤的影响及其机制。方法 将雄性C57小鼠随机分为正常组(control组)、慢性间歇性低氧组(CIH组)和SAHA干预组(CIH+SAHA组),将CIH+SAHA组与CIH组小鼠放入低氧仓内进行间歇性低氧处理,每天8 h,持续4周,从第3周起,每日造模前给予实验小鼠腹腔注射SAHA 50 mg/(kg·d),给药持续2周。实验第4周后,测量小鼠体质量,然后处死小鼠,HE染色观察肝组织形态学变化,测定血清中谷丙转氨酶(ALT)和谷草转氨酶(AST)水平,Western blot检测小鼠肝组织中裂解型胱天蛋白酶3(c-Caspase-3)和Caspase-9的蛋白水平,TUNEL试剂盒检测小鼠肝组织细胞凋亡情况。结果 与control组相比,间歇性低氧4周后,CIH组小鼠肝指数显著升高且肝组织损伤明显,肝组织细胞凋亡相关蛋白Caspase-9和c-Caspase-3的表达水平显著增高,肝组织细胞凋亡比例增加(P<0.05);与CIH组相比,CIH+SAHA组肝指数、ALT、AST水平以及Caspase-9和c-Caspase-3蛋白表达水平、肝组织细胞凋亡指标降低(P<0.05)。结论 SAHA可能通过抑制凋亡相关蛋白Caspase-9/Caspase-3,进而改善慢性间歇性低氧引起的小鼠肝损伤。

新型Bcl-2小分子抑制剂的设计、合成与初步活性评价

目的设计并合成一系列新型Bcl-2小分子抑制剂,测试其对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用的抑制活性,初步探究其构效关系,为后续相关研究提供参考。方法以临床化合物BGB-11417为先导化合物,通过骨架跃迁等方法,设计新型含螺环结构的Bcl-2小分子抑制剂。以苯甲醛为原料,通过取代、环化和偶联等反应合成目标化合物,并通过1H NMR和LC-MS进行结构确定。采用时间分辨荧光共振能量转移(TR-FRET)评价目标化合物对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用的抑制能力。结果共合成8个新型螺环Bcl-2小分子抑制剂,其中29a[IC_(50)(Bcl-2):0.8 nmol·L^(-1),IC_(50)(Bcl-2 G101V):55.41 nmol·L^(-1)],29d[IC_(50)(Bcl-2):0.27 nmol·L^(-1),IC_(50)(Bcl-2 G101V):18.65 nmol·L^(-1)]对Bcl-2和Bcl-2 G101V与BH3-only蛋白的相互作用有较好的抑制活性。结论建立了一种新型螺环Bcl-2小分子抑制剂合成方法,发现了对Bcl-2和Bcl-2 G101V(Bcl-2突变体)与BH3-only蛋白相互作用有较好抑制活性的新化合物,其中29d具有进一步研究的价值。

肝外胆管癌ANXA1和XIAP表达的临床病理研究

目的探讨膜联蛋白A1(ANXA1)和X连锁凋亡抑制蛋白(XIAP)表达与肝外胆管癌临床病理特征及预后的关系。方法58例肝外胆管癌手术切除标本及30例癌旁胆管组织,应用EliVision^(TM)plus免疫组织化学染色二步法检测ANXA1和XIAP在癌组织及癌旁胆管组织中的表达,分析ANXA1和XIAP与肝外胆管癌患者性别、年龄、癌灶大小、分化等级、TNM分期、浸润转移及3年生存率7项临床病理特征的相关性。结果癌组织ANXA1和XIAP阳性率分别为84.5%(49/58)和81.0%(47/58),癌旁胆管组织分别为30.0%(9/30)和26.7%(8/30),差异有统计学意义(均P<0.05)。ANXA1表达与性别、年龄、分化等级无关(均P>0.05),在癌灶>1.5 cm、TNM(Ⅲ+Ⅳ)期、有浸润转移和3年内死亡患者中,其阳性率显著高于癌灶≤1.5 cm、TNM(Ⅰ+Ⅱ)期、无浸润转移和3年内生存患者(均P<0.05);XIAP表达与性别、年龄无关(均P>0.05),在癌灶>1.5 cm、低分化、TNM(Ⅲ+Ⅳ)期、有浸润转移和3年内死亡患者中,其阳性率显著高于癌灶≤1.5 cm、(高+中)分化、TNM(Ⅰ+Ⅱ)期、无浸润转移和3年内生存患者(均P<0.05)。ANXA1和XIAP表达呈正相关性(r=0.54)。结论ANXA1和XIAP与肝外胆管癌发生发展显著相关,两者表达上调预示癌灶大、分期晚、易浸润转移、生存期短。

X-染色体连锁凋亡抑制蛋白相关因子1通过自噬途径调控Caspase 1介导的胃肠道间质瘤细胞焦亡

目的:探究X-染色体连锁凋亡抑制蛋白相关因子1(XAF1)对人胃肠道间质瘤(GIST)细胞中半胱氨酸蛋白酶-1(Caspase 1)介导的细胞焦亡的影响及机制。方法:实时荧光定量PCR和Western blot检测人正常胃黏膜上皮细胞系(RGM-1)和GIST细胞系(GIST-T1、GIST-430、GIST-882)中XAF1的表达水平;采用脂质体介导转染法将pcDNA3.1-XAF1重组质粒染进GIST-882细胞,以构建高表达XAF1的GIST-882细胞。GIST-882细胞分为对照组、pcDNA3.1组(转染pcDNA3.1空载体质粒)、pcDNA3.1-XAF1组(转染pcDNA3.1-XAF1重组质粒)、pcDNA3.1-XAF1+Rap组(转染pcDNA3.1-XAF1重组质粒并给予自噬激活剂雷帕霉素处理),免疫荧光染色检测各组细胞内微管相关蛋白1轻链3(LC3)表达情况,Western blot检测各组细胞中LC3Ⅱ/LC3Ⅰ比值与Beclin1蛋白表达水平,ELISA法检测各组细胞上清中白细胞介素,IL-1β、IL-18含量,乳酸脱氢酶(LDH)释放实验检测各组细胞LDH释放率,Western blot检测各组细胞中Caspase-1及其活化形式cleaved-Caspase-1、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、消皮素D(GSDMD)蛋白表达水平。结果:与RGM-1细胞比较,GIST-T1、GIST-430、GIST-882细胞中XAF1 mRNA和蛋白相对表达量显著减少(P<0.05);细胞转染后,pcDNA3.1-XAF1组GIST-882细胞中XAF1 mRNA和蛋白相对表达量均显著高于pcDNA3.1组、对照组(P<0.05)。与对照组比较,pcDNA3.1-XAF1组GIST-882细胞内LC3荧光强度明显减弱,LC3Ⅱ/LC3Ⅰ比值、Beclin1蛋白相对表达量显著减少(P<0.05),细胞上清液IL-1β、IL-18含量及LDH释放率显著升高(P<0.05),Caspase-1、cleaved-Caspase-1、NLRP3、GSDMD蛋白相对表达量也显著增加(P<0.05);与pcDNA3.1-XAF1组比较,pcDNA3.1-XAF1+Rap组GIST-882细胞内LC3荧光强度明显增强,LC3Ⅱ/LC3Ⅰ比值、Beclin1蛋白相对表达量显著增加(P<0.05),同时,细胞上清液IL-1β、IL-18含量及LDH释放率显著下降(P<0.05),Caspase-1、cleaved-Caspase-1、NLRP3、GSDMD蛋白相对表达量也显著减少(P<0.05)。结论:XAF1在GIST细胞中表达下降,提高XAF1表达能通过抑制自噬从而促进Caspase 1介导的GIST细胞焦亡,发挥抗肿瘤作用。

硫氧还蛋白还原酶结构与肿瘤治疗的研究进展

硫氧还蛋白还原酶(thioredoxin reductase,TrxR)是硫氧还蛋白(thioredoxin,Trx)系统里主要的功能蛋白,在调节多种细胞氧化还原信号通路中起关键作用。近年来,TrxR/Trx被越来越多地认为是肿瘤发展的重要调节剂,因此靶向TrxR/Trx是一种很有前景的肿瘤治疗策略。该文对TrxR的结构特点、与肿瘤相关的生理功能、TrxR抑制剂进行综述,以对TrxR的研究提供参考。

Mitochondrial division inhibitor 1 protects cortical neurons from excitotoxicity:a mechanistic pathway

Mitochondrial division inhibitor 1（Mdivi-1） is a selective cell-permeable inhibitor of dynamin-related protein-1（Drp1） and mitochondrial division.To investigate the effect of Mdivi-1 on cells treated with glutamate,cerebral cortex neurons isolated from neonatal rats were treated with 10 m M glutamate for 24 hours.Normal cultured cells and dimethyl sulfoxide-cultured cells were considered as controls.Apoptotic cells were detected by flow cytometry.Changes in mitochondrial morphology were examined by electron microscopy.Drp1,Bax,and casp ase-3 expression was evaluated by western blot assays and immunocytochemistry.Mitochondrial membrane potential was detected using the JC-1 probe.Twenty-four hours after 10 m M glutamate treatment,Drp1,Bax and caspase-3 expression was upregulated,Drp1 and Bax were translocated to mitochondria,mitochondrial membrane potential was decreased and the rate of apoptosis was increased.These effects were inhibited by treatment with 50 μM Mdivi-1 for 2 hours.This finding indicates that Mdivi-1 is a candidate neuroprotective drug that can potentially mitigate against neuronal injury caused by glutamate-induced excitotoxicity.

喉癌患者细胞凋亡抑制蛋白1的表达情况及预后影响因素

目的探讨喉癌患者喉癌组织细胞凋亡抑制蛋白1(c-IAP1)的表达情况及影响患者预后的因素。方法回顾性分析60例喉癌患者的临床资料,采集患者术后喉癌组织与癌旁组织病理切片,采用免疫组织化学法测定c-IAP1的表达情况。比较喉癌患者喉癌组织与癌旁组织中c-IAP1的表达情况,以及不同预后患者喉癌组织中c-IAP1的表达情况。采用多因素COX回归模型分析影响喉癌患者预后的因素。结果喉癌组织中c-IAP1高表达率高于癌旁组织(P<0.05)。本组60例患者中,预后不良42例(70.00%),预后良好18例(30.00%),预后不良患者喉癌组织c-IAP1高表达率高于预后良好患者(P<0.05)。多因素COX回归分析结果显示,c-IAP1表达情况、组织学分级、肿瘤大小、淋巴结转移情况、临床分期是喉癌患者预后的影响因素(均P<0.05)。结论喉癌患者喉癌组织中c-IAP1阳性表达率升高,c-IAP1表达情况、组织学分级、肿瘤大小、淋巴结转移情况、临床分期是喉癌患者预后的影响因素。