Aldo-keto reductase family member C3(AKR1C3)promotes hepatocellular carcinoma cell growth by producing prostaglandin F2α

Hepatocellular carcinoma(HCC)is a leading cause of death worldwide.Current therapies are effective for HCC patients with early disease,but many patients suffer recurrence after surgery and have a poor response to chemotherapy.Therefore,new therapeutic targets are needed.We analyzed gene expression profiles between HCC tissues and normal adjacent tissues from public databases and found that the expression of genes involved in lipid metabolism was significantly different.The analysis showed that AKR1C3 was upregulated in tumors,and high AKR1C3 expression was associated with a poorer prognosis in HCC patients.In vitro,assays demonstrated that the knockdown of AKR1C3 or the addition of the AKR1C3 inhibitor indomethacin suppressed the growth and colony formation of HCC cell lines.Knockdown of AKR1C3 in Huh7 cells reduced tumor growth in vivo.To explore the mechanism,we performed pathway enrichment analysis,and the results linked the expression of AKR1C3 with prostaglandin F2 alpha(PGF2a)downstream target genes.Suppression of AKR1C3 activity reduced the production of PGF2a,and supplementation with PGF2a restored the growth of indomethacin-treated Huh7 cells.Knockdown of the PGF receptor(PTGFR)and treatment with a PTGFR inhibitor significantly reduced HCC growth.We showed that indomethacin potentiated the sensitivity of Huh7 cells to sorafenib.In summary,our results indicate that AKR1C3 upregulation may promote HCC growth by promoting the production of PGF2α,and suppression of PTGFR limited HCC growth.Therefore,targeting the AKR1C3-PGF2a-PTGFR axis may be a new strategy for the treatment of HCC.

MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1-extracellular signal-regulated kinase-1/2 axis

BACKGROUND MicroRNAs(miRNAs)regulate gene expression and play a critical role in cancer physiology.However,there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer(GC).AIM To investigate the role and molecular mechanism of miRNA-145-5p(miR145-5p)in the progression of GC.METHODS Real-time polymerase chain reaction(RT-PCR)was used to detect miRNA expression in human GC tissues and cells.The ability of cancer cells to migrate and invade was assessed using wound-healing and transwell assays,respectively.Cell proliferation was measured using cell counting kit-8 and colony formation assays,and apoptosis was evaluated using flow cytometry.Expression of the epithelial-mesenchymal transition(EMT)-associated protein was determined by Western blot.Targets of miR-145-5p were predicated using bioinformatics analysis and verified using a dual-luciferase reporter system.Serpin family E member 1(SERPINE1)expression in GC tissues and cells was evaluated using RT-PCR and immunohistochemical staining.The correlation between SERPINE1 expression and overall patient survival was determined using Kaplan-Meier plot analysis.The association between SERPINE1 and GC progression was also tested.A rescue experiment of SERPINE1 overexpression was conducted to verify the relationship between this protein and miR-145-5p.The mechanism by which miR-145-5p influences GC progression was further explored by assessing tumor formation in nude mice.RESULTS GC tissues and cells had reduced miR-145-5p expression and SERPINE1 was identified as a direct target of this miRNA.Overexpression of miR-145-5p was associated with decreased GC cell proliferation,invasion,migration,and EMT,and these effects were reversed by forcing SERPINE1 expression.Kaplan-Meier plot analysis revealed that patients with higher SERPINE1 expression had a shorter survival rate than those with lower SERPINE1 expression.Nude mouse tumorigenesis experiments confirmed that miR-145-5p targets SERPINE1 to regulate extracellular signal-regulated kinase-1/2(ERK1/2).CONCLUSION This study found that miR-145-5p inhibits tumor progression and is expressed in lower amounts in patients with GC.MiR-145-5p was found to affect GC cell proliferation,migration,and invasion by negatively regulating SERPINE1 levels and controlling the ERK1/2 pathway.

自然杀伤细胞2组成员A、程序性死亡因子配体1表达检测对PD-L1阻断免疫治疗肌层浸润性膀胱癌反应性的预测价值

目的:探究自然杀伤细胞2组成员A(NKG2A)及程序性死亡因子配体1(PD-L1)表达检测对PD-L1阻断免疫治疗肌层浸润性膀胱癌反应性的预测价值。方法:选取100例肌层浸润性膀胱癌患者作为研究对象,对其行PD-L1阻断免疫治疗后,根据治疗反应性将其分为缓解组和未缓解组;对缓解组患者随访3个月,将其分为复发组和未复发组。比较两组患者NKG2A和PD-L1在CD4^(+)和CD8^(+)上的表达水平,采用ROC曲线分析NKG2A和PD-L1预测膀胱癌患者治疗反应性的价值。结果:100例研究对象中,缓解组患者72例(72.00%),未缓解组患者28例(28.00%),两组性别、年龄比较无统计学差异(均P>0.05),但缓解组患者肿瘤直径小于未缓解组,NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达水平均低于未缓解组(均P<0.05)。膀胱癌患者NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达预测免疫治疗后缓解的AUC值分别为0.771、0.724、0.710;联合诊断的AUC为0.836。72例缓解患者中,出现复发29例(40.28%),未出现复发43例(59.72%),两组性别、年龄以及肿瘤直径比较无统计学差异(均P>0.05),但复发组NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达水平均高于未复发组(均P<0.05)。NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达预测膀胱癌患者缓解后复发的AUC值分别为0.775、0.740、0.728;联合诊断的AUC为0.874。结论:NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)在不同治疗反应性肌层浸润性膀胱癌患者外周血中表达水平不同,三者对于膀胱癌患者PD-L1阻断免疫治疗反应性均有一定的预测价值,且三者联合预测效能最佳。

PIM1基因对急性髓系白血病U937细胞增殖、凋亡及JAK2/STAT3信号通路的影响

目的:探讨PIM1基因对急性髓系白血病(AML)U937细胞增殖、凋亡的影响,以及对JAK2/STAT3通路的调控作用。方法:收集初诊成人AML患者和单纯缺铁性贫血患者的骨髓单个核细胞,荧光定量PCR检测PIM1 mRNA表达。将AML细胞系U937细胞分为:U937组(U937细胞正常培养)、Si-PIM1组(U937细胞转染含PIM1 mRNA的低表达腺病毒载体)、Si-NC组(U937细胞转染不含PIM1 mRNA的低表达腺病毒载体)、CoA1组(U937细胞中加入浓度为20μmol/L的JAK2激活剂CoA1)、Si-PIM1+CoA1组(U937细胞转染含PIM1 mRNA低表达的腺病毒载体并加入浓度为20μmol/L的CoA1)。培养24 h。荧光定量PCR和蛋白印迹法检测U937细胞PIM1 mRNA和蛋白、JAK2/STAT3通路、细胞周期、凋亡相关蛋白表达;噻唑蓝法检测细胞增殖活性;流式细胞术检测细胞周期变化及凋亡率。结果:AML患者骨髓单个核细胞中PIM1 mRNA表达水平高于单纯缺铁性贫血患者(P<0.05)。与U937组相比,Si-PIM1组细胞PIM1 mRNA和蛋白、p-JAK2/JAK2、p-STAT3/STAT3、Cyclin D1、CDK2蛋白、细胞增殖活性、S期比例、G2/M期比例降低(均P<0.05),p27、Caspase-3蛋白、G0/G1期、凋亡率升高(均P<0.05),而CoA1组上述指标的变化情况与Si-PIM1组正好相反,CoA1可逆转Si-PIM1对U937细胞的作用效果。U937组、Si-PIM1+CoA1组、Si-NC组U937细胞上述指标差异无统计学意义(P>0.05)。结论:敲低PIM1基因表达可抑制U937细胞增殖、促进凋亡,缓解ALM进程,且上述作用可能与抑制JAK2/STAT3通路活化有关。

结直肠癌组织中NR2F1的表达及其与预后的相关性

目的检测结直肠癌中核受体亚家族2组F成员1(NR2F1)的蛋白表达水平,并探讨其与患者预后的关系。方法选择2012年8月至2013年9月在苏州市立医院进行根治性切除术的56例结直肠癌患者作为研究对象,采集患者的结直肠癌组织标本及对应的癌旁组织标本。采用EnVision法测定结直肠癌组织和癌旁组织的NR2F1的蛋白表达水平,比较不同NR2F1蛋白表达水平结直肠癌患者的临床病理特征。采用Kaplan-Meier法绘制结直肠癌患者的生存曲线,分析NR2F1蛋白表达水平与结直肠癌的预后的相关性。结果结直肠癌组织的NR2F1高表达率为48.21%,明显高于癌旁组织的7.14%,差异有统计学意义(P<0.05);NR2F1高表达者的无脉管侵犯率为85.19%,明显高于NR2F1低表达者的58.62%,差异有统计学意义(P<0.05);NR2F1高表达患者突破浆膜层的占比为48.15%,明显低于NR2F1低表达者的65.52%,且随着浸润深度从固有肌层到突破浆膜层,NR2F1低表达增加,差异均具有统计学意义(P<0.05);NR2F1高表达和低表达患者在性别、年龄、肿块大小、分化程度、淋巴结转移方面比较,差异均无统计学意义(P>0.05);经Kaplan-Meier生存曲线分析结果显示,结直肠癌的NR2F1高表达患者的10年总生存率为26.79%,明显高于NR2F1低表达患者的17.86%,差异有统计学意义(P<0.05)。结论结直肠癌的NR2F1表达升高,NR2F1可能参与了结直肠癌的发生、侵袭,对患者的预后造成一定的影响。

三磷酸腺苷结合盒转运蛋白G2与痛风病关联的病理机制研究进展

痛风病是指与高尿酸血症、尿酸盐沉积(关节、肾脏)等尿酸代谢紊乱密切相关的进展性代谢疾病,根据其病理进展特点和临床表现,其病程可分为高尿酸血症阶段、尿酸盐沉积阶段、痛风性炎症阶段。三磷酸腺苷结合盒转运蛋白G2(ABCG2)是高通量的尿酸转运体,既参与了机体尿酸外排的肠-肾途径,又与痛风性炎症密切相关。通过切入ABCG2,对痛风病病理机制研究进展进行综述,以期为临床痛风病的深入研究及防治提供新的见解。

垂体泌乳素瘤患者组织NR2C2、EMMPRIN水平变化及其与肿瘤侵袭性、临床预后的关系探讨

目的探讨垂体泌乳素瘤组织中核受体亚家族2c成员2(NR2C2)和细胞外基质金属蛋白酶诱导因子(EMMPRIN)的水平变化及其与肿瘤侵袭性、临床预后的关系。方法选择2019年6月—2021年6月本院收治的78例垂体泌乳素瘤患者为研究对象,患者均行手术切除,术后随访时间1年。采用免疫组化法检测NR2C2和EMMPRIN表达情况,对比癌组织、癌旁组织中NR2C2和EMMPRIN表达,分析垂体泌乳素瘤患者癌组织中NR2C2和EMMPRIN表达情况与临床病理参数的关系,分析影响垂体泌乳素瘤患者预后的因素。结果癌组织中NR2C2阳性表达率高于癌旁组织(P<0.05),EMMPRIN阳性表达率低于癌旁组织(P<0.05),NR2C2、EMMPRIN表达均是预后不良的独立危险因素(P<0.05);受试者工作特征曲线(ROC)分析显示,垂体泌乳素瘤组织NR2C2、EMMPRIN两者联合预测垂体泌乳素瘤患者预后不良的曲线下面积(AUC)为0.886,高于NR2C2、EMMPRIN单独预测的AUC(Z=5.245,P=0.014;Z=3.652,P=0.028)。结论垂体泌乳素瘤患者癌组织中NR2C2和EMMPRIN表达与肿瘤侵袭性、临床预后有关,NR2C2阳性患者预后不良风险高。

鄂尔多斯盆地铁边城地区长2油层组剩余油模拟

为了实现油田增储上产及老井挖潜的目标,提升剩余油资源开发潜力,通过岩心、薄片、电镜、物性、模拟方法系统的研究了鄂尔多斯盆地西北部姬塬油田铁边城地区长2段主力油层的剩余油分布特征。研究显示,长2_(1)^(3-2)和长2_(1)^(3-3)小层为主力产油层,平均厚度分别为12.2 m及11.1 m。目的层砂岩储层的孔隙度通常介于12%~18%,渗透率则通常低于8 mD。根据相控原则,建立了主力产层的物理属性模型及流体模型;进而,开展了剩余油分布数值模拟研究。通过基于实测产水及产油量约束的历史生产曲线拟合,获得了剩余油的纵横向分布结果。研究显示,剩余油分布受砂体边界、物性差异及注采关系的影响。剩余油整体上成片分布;但是,受油藏物性差异的影响,注水多呈不均匀受效,因而造成局部剩余油富集。剩余油主要分布在砂体边界处,这些区域无井网控制,因而水驱程度低,剩余油分布较多。受长期注采的影响,井间会形成高渗区,这些其余类似高速通道,会发生快速水淹,其周边则形成剩余油滞留区。可见,剩余油富集明显受微相、物性、注水及井网因素的控制。

雄黄主要成分As_(2)S_(2)通过“ANP32A-INHAT-H3乙酰化”轴抑制三阴性乳腺癌细胞增殖、迁移的表观遗传调控机制

目的探究雄黄主要成分二硫化二砷(As_(2)S_(2))对三阴性乳腺癌(triple negative breast cancer,TNBC)的作用及表观遗传调控机制。方法采用CCK-8、平板克隆形成和细胞划痕实验探究As_(2)S_(2)对人正常乳腺上皮细胞MCF-10A及TNBC细胞增殖和迁移的影响;4D-label free定量蛋白质组学分析挖掘As_(2)S_(2)抗TNBC的潜在干预靶点酸性核磷蛋白家族成员32A(acidic nuclear phosphoprotein family member 32A,ANP32A);慢病毒感染法构建ANP32A过表达敲低细胞株,探究潜在靶点ANP32A对As_(2)S_(2)抗TNBC作用的影响;蛋白质免疫共沉淀和Western blot实验探究As_(2)S_(2)是否通过ANP32A调控TNBC细胞H3乙酰化。结果As_(2)S_(2)对人正常乳腺上皮细胞MCF-10A影响甚微,但显著抑制TNBC细胞增殖和迁移,且呈剂量依赖性;4D-lable free定量蛋白质组学分析结果显示,促癌因子ANP32A被As_(2)S_(2)显著下调,且ANP32A表达影响As_(2)S_(2)在TNBC中的抗增殖和迁移效果。As_(2)S_(2)能下调ANP32A的蛋白水平,抑制乙酰转移酶抑制剂复合物亚基的招募,增加H3乙酰化水平。结论As_(2)S_(2)通过下调ANP32A蛋白调控TNBC细胞中H3乙酰化,抑制TNBC细胞增殖和迁移。

CYP2D6基因分型影响因素研究进展

细胞色素P450家族2亚科D成员6(CYP2D6)是细胞色素P450酶家族中的重要药物代谢酶,是抗抑郁药物、抗精神病药物和阿片类镇痛药物等主要的代谢酶。CYP2D6基因位点的复杂性和诸多等位基因突变体造成了CYP2D6表型的多态性,目前已报道170余种等位基因突变体。CYP2D6酶活性变化很大,从无活性到超快代谢均存在,根据酶活性可将不同表型携带者分为超快代谢者、正常代谢者、中间代谢者和弱代谢者。随着个体化医疗的发展,CYP2D6基因分型试验可以辅助药物遗传学和基因分型技术的研究和临床应用。然而,由于CYP2D6基因存在复杂的突变,包括单核苷酸突变、插入、缺失、基因拷贝数变异和基因重组。CYP2D6基因不仅存在个体化差异,且在不同种族之间等位基因的频率也明显不同。另外,人体内同时存在与CYP2D6同源性很高的非功能性基因CYP2D7,通过基因检测分析CYP2D6表型是一项非常具有挑战性的工作。文章总结了CYP2D6基因的多态性和基因分型的复杂性,并分析了部分不同的基因型突变对CYP2D6基因分型的影响,以帮助临床通过基因分型方法对CYP2D6酶活性进行预测。

茯苓酸通过调节Nrf2/SLC7A11/GPX4信号通路抑制氧糖剥夺/复氧诱导的心肌细胞铁死亡

目的:探究茯苓酸通过调节核因子E2相关因子2(Nrf2)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)信号通路抑制氧糖剥夺/复氧(OGD/R)诱导的心肌细胞铁死亡的机制。方法:体外培养大鼠心肌细胞H9c2,诱导建立细胞OGD/R模型后以0、1.0、2.5、5.0、10.0、20.0μmol/L茯苓酸处理24 h,通过细胞计数试剂盒-8(CCK-8)法检测各处理组细胞活力后筛选出合适的茯苓酸作用浓度。将H9c2细胞随机分为对照组、模型组、茯苓酸组、ML385组(Nrf2抑制剂组)、茯苓酸+ML385组,除对照组外其余各组建立细胞OGD/R模型后以茯苓酸、ML385分组处理,采用CCK-8法与流式细胞实验分别检测各组H9c2细胞活力、凋亡率;采用试剂盒检测各组H9c2细胞培养基上清中乳酸脱氢酶(LDH)、促炎因子[前列腺素E2(PGE2)、肿瘤坏死因子-α(TNF-α)]、抗炎因子白细胞介素(IL)-10水平及细胞抗氧化因子[过氧化氢酶(CAT)、谷胱甘肽(GSH)]、铁死亡相关指标[铁含量、丙二醛(MDA)]水平;采用免疫印迹实验检测各组H9c2细胞凋亡及Nrf2/SLC7A11/GPX4信号通路相关蛋白表达。结果:与对照组比较,模型组细胞凋亡率、LDH释放量、细胞培养基上清中PGE2及TNF-α水平、细胞铁含量、MDA水平及Bax、Caspase-3蛋白表达均升高(P<0.05),细胞活力、细胞培养基上清中IL-10水平、细胞CAT及GSH水平、Bcl-2及Nrf2、SLC7A11、GPX4蛋白表达降低(P<0.05)。与模型组比较,茯苓酸组细胞凋亡率、LDH释放量、细胞培养基上清中PGE2及TNF-α水平、细胞铁含量、MDA水平及Bax、Caspase-3蛋白表达均降低(P<0.05),细胞活力、细胞培养基上清中IL-10水平、细胞CAT及GSH水平、Bcl-2及Nrf2、SLC7A11、GPX4蛋白表达升高(P<0.05);ML385组细胞凋亡率、LDH释放量、细胞培养基上清中PGE2及TNF-α水平、细胞铁含量、MDA水平及Bax、Caspase-3蛋白表达均升高(P<0.05),细胞活力、细胞培养基上清中IL-10水平、细胞CAT及GSH水平、Bcl-2及Nrf2、SLC7A11、GPX4蛋白表达降低(P<0.05)。与茯苓酸组比较,茯苓酸+ML385组细胞凋亡率、LDH释放量、细胞培养基上清中PGE2及TNF-α水平、细胞铁含量、MDA水平及Bax、Caspase-3蛋白表达均升高(P<0.05),细胞活力、细胞培养基上清中IL-10水平、细胞CAT及GSH水平、Bcl-2及Nrf2、SLC7A11、GPX4蛋白表达降低(P<0.05)。结论:茯苓酸可通过激活Nrf2/SLC7A11/GPX4信号而抑制OGD/R诱导的心肌细胞炎症、脂质过氧化与铁死亡,增强其抗氧化活性及细胞活力,最终减轻其细胞凋亡损伤。

hsa_circ_0000520通过调控miR-556-5p/SLC38A2促进乳腺癌的发生和转移

目的:探究hsa_circ_0000520促进乳腺癌发生和转移的作用机制。方法:双荧光素酶报告基因实验、RNA pull-down实验验证miR-556-5p与hsa_circ_0000520、溶质载体家族38成员2(SLC38A2)的靶向关系。MCF7细胞分为sh-NC组、sh-hsa_circ_0000520组、sh-hsa_circ_0000520+anti-NC组、sh-hsa_circ_0000520+anti-miR-556-5p组,qRT-PCR或Western blot检测细胞中hsa_circ_0000520、miR-556-5p、SLC38A2表达水平;MTT检测、平板克隆形成实验评估细胞增殖能力;划痕愈合实验、Transwell实验评估细胞迁移、侵袭能力。通过裸鼠成瘤实验评估hsa_circ_0000520对miR-556-5p/SLC38A2的调控作用及对移植瘤生长的影响。结果:经验证,MCF7细胞中miR-556-5p与hsa_circ_0000520、SLC38A2均存在靶向关系。与sh-NC组比较,sh-hsa_circ_0000520组可降低MCF7细胞中hsa_circ_0000520、SLC38A2 mRNA和蛋白表达水平、细胞活力、克隆形成数目、划痕愈合率及迁移、侵袭细胞数(P<0.05),升高miR-556-5p表达水平(P<0.05);与sh-hsa_circ_0000520+anti-NC组比较,sh-hsa_circ_0000520+anti-miR-556-5p组可降低MCF7细胞中miR-556-5p表达水平(P<0.05),升高SLC38A2 mRNA和蛋白表达水平、细胞活力、克隆形成数目、划痕愈合率及迁移、侵袭细胞数(P<0.05),而对hsa_circ_0000520表达无显著影响(P>0.05)。裸鼠成瘤实验结果表明,敲低移植瘤中hsa_circ_0000520的表达可升高miR-556-5p表达水平并降低SLC38A2 mRNA和蛋白表达水平,同时降低肿瘤体积和肿瘤重量(P<0.05)。结论:hsa_circ_0000520可能通过靶向调控miR-556-5p/SLC38A2促进乳腺癌的发生和转移。

嗅觉受体家族13亚家族C成员2在肝细胞癌中的表达及临床病理意义

目的探讨嗅觉受体家族13亚家族C成员2(olfactory receptor family 13 subfamily C member 2,OR13C2)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及临床病理意义。方法收集2017年8月至2022年3月于右江民族医学院附属医院进行手术切除治疗的HCC患者为研究对象,取石蜡包埋的癌组织及癌旁非癌肝组织适量,采用免疫组化染色EnVision二步法检测OR13C2在上述组织中的表达情况,通过Logistic回归模型分析OR13C2的表达与HCC组织学分级、癌旁肝纤维化等临床病理特征的相关性。结果OR13C2在HCC癌组织中的表达明显低于癌旁肝组织,差异具有统计学意义(P<0.05);HCC肿瘤组织中OR13C2的表达与HCC组织学分级差异有统计学意义(P<0.05),HCC中OR13C2的表达与HCC的组织学分级呈负相关(r=-0.213,P<0.05),OR13C2表达下调为肿瘤去分化风险因子(风险值为3.914,P<0.05);OR13C2在癌组织中的表达与HCC的发病年龄、性别、包膜侵犯、临床分期、乙型肝炎病毒感染、术前血清甲胎蛋白及微血管侵犯以及癌组织中Glypican-3、Arginase-1表达的差异均无统计学意义(P>0.05),OR13C2在癌旁肝组织中的表达与肝纤维化分期、慢性肝炎分级差异无统计学意义(P>0.05)。结论HCC癌组织中OR13C2表达水平下调,这种下调与肿瘤的组织学分级负相关,是一个肿瘤去分化的风险因子。这些结果提示OR13C2表达失调在HCC发生中可能起一定的促进作用。

miR-144-3p通过调控ABCG2信号通路对膀胱癌细胞的影响

目的探究miR-144-3p通过调控ATP结合转运蛋白G家族成员2(ATP-binding cassette superfamily G member 2,ABCG2)信号通路对膀胱癌细胞影响。方法选取2023年5月黑龙江省医院泌尿科实验室1株人膀胱癌T24细胞株为研究对象,细胞培养后接种6孔板中,将其随机分为A组、B组、C组,每组细胞均设置3个平行样本,每个样本细胞检测均设置3个复孔(每组共计12个样本量)。A组转染miR-144-3p模拟物,B组转染miR-144-3p抑制剂,C组转染无意义序列,另选正常膀胱上皮SV-HUC-1细胞株为D组,以实时定量联合酶链式反应(quantification real-time polymerase chian reaction,qRT-PCR)法检测4组细胞中miR-144-3p表达情况,以CCK-8法检测4组细胞增殖活力,以Transwell检测细胞迁移数量,检测ABCG2通路蛋白表达情况;以双荧光素酶报告基因实验预测miR-144-3p与ABCG2靶向结合关系。结果转染前,T24细胞中miR-144-3p表达水平较SV-HUC-1细胞低,ABCG2水平较SV-HUC-1细胞高,差异有统计学意义(t=8.145、13.928,P<0.05);转染48 h时,miR-144-3p水平由低至高依次为B组、C组、A组,差异有统计学意义(P<0.05),ABCG2表达水平由低至高依次为A组、C组、B组,差异有统计学意义(P<0.05);转染48 h后,细胞增殖能力(A值)由低至高依次为A组、C组、B组,细胞凋亡率由低至高依次为B组、C组、A组,细胞迁移能力(个数)由低至高依次为A组、C组、B组,ABCG2表达水平由低至高依次为A组、C组、B组,差异有统计学意义(P<0.05);经在线数据库TargetScan预测,测得ABCG23’UTR与miR-144-3p存在结合靶点;共转染48 h时检测,3’-UTR-WT miR-144-3p组荧光素酶活性较3’-UTR-Wut miR-144-3p组低,差异有统计学意义(P<0.05);3’-UTA-WT NC组、3’-UTR-Mut NC组荧光素酶活性相近,差异无统计学意义(P>0.05)。结论ABCG23’UTR与miR-144-3p存在结合靶点,miR-144-3p表达上调可抑制ABCG2表达,并抑制细胞迁移、增殖,促进细胞凋亡。

Geological characteristics and accumulation mechanisms of the "continuous" tight gas reservoirs of the Xu2 Member in the middle-south transition region,Sichuan Basin,China

＂Continuous＂ tight gas reservoirs are those reservoirs which develop in widespread tight sandstones with a continuous distribution of natural gas. In this paper, we summarize the geological features of the source rocks and ＂＇continuous＂ tight gas reservoirs in the Xujiahe Formation of the middle- south transition region, Sichuan Basin. The source rocks of the Xul Member and reservoir rocks of the Xu2 Member are thick （Xul Member： 40 m, Xu2 Member： 120 m） and are distributed continuously in this study area. The results of drilled wells show that the widespread sandstone reservoirs of the Xu2 Member are charged with natural gas. Therefore, the natural gas reservoirs of the Xu2 Member in the middle-south transition region are ＂continuous＂ tight gas reservoirs. The accumulation of ＂continuous＂ tight gas reservoirs is controlled by an adequate driving force of the pressure differences between source rocks and reservoirs, which is demonstrated by a ＂one-dimensional＂ physical simulation experiment. In this simulation, the natural gas of＂continuous＂ tight gas reservoirs moves tbrward with no preferential petroleum migration pathways （PPMP）, and the natural gas saturation of＂continuous＂ tight gas reservoirs is higher than that of conventional reservoirs.

致密碳酸盐岩储层微观结构特征及发育影响因素——以鄂尔多斯盆地东部马五_(1+2)亚段为例

为了明确鄂尔多斯盆地东部马五_(1+2)亚段致密碳酸盐岩储层发育有利区,应用铸体薄片鉴定、高压压汞、低温液氮吸附及CT扫描的方法,对研究区马五_(1+2)亚段致密碳酸盐岩储层特征进行了系统研究。结果表明:研究区储层岩性以含硬石膏结核白云岩、角砾白云岩(渗流粉砂充填)、砂屑白云岩和细粉晶白云岩为主,其中,含硬石膏结核白云岩为优质储集岩,但其发育程度及规模较有限;角砾白云岩(渗流粉砂充填)储集性能略差,但其分布规模大,更具勘探潜力。主要受沉积环境、成岩作用及不整合3类因素的影响,研究区碳酸盐岩储层与靖边气田相比,角砾化严重,含硬石膏结核白云岩发育厚度明显变小,而且更为致密。结合有利储层的分布特征,优选了榆林—米脂地区为下一步勘探的重点目标区。

Solute carrier family 2 members 1 and 2 as prognostic biomarkers in hepatocellular carcinoma associated with immune infiltration

BACKGROUND Metabolic reprogramming has been identified as a core hallmark of cancer.Solute carrier family 2 is a major glucose carrier family.It consists of 14 members,and we mainly study solute carrier family 2 member 1(SLC2A1)and solute carrier family 2 member 2(SLC2A2)here.SLC2A1,mainly existing in human erythrocytes,brain endothelial cells,and normal placenta,was found to be increased in hepatocellular carcinoma(HCC),while SLC2A2,the major transporter of the normal liver,was decreased in HCC.AIM To identify if SLC2A1 and SLC2A2 were associated with immune infiltration in addition to participating in the metabolic reprogramming in HCC.METHODS The expression levels of SLC2A1 and SLC2A2 were tested in HepG2 cells,HepG215 cells,and multiple databases.The clinical characteristics and survival data of SLC2A1 and SLC2A2 were examined by multiple databases.The correlation between SLC2A1 and SLC2A2 was analyzed by multiple databases.The functions and pathways in which SLC2A1,SLC2A2,and frequently altered neighbor genes were involved were discussed in String.Immune infiltration levels and immune marker genes associated with SLC2A1 and SLC2A2 were discussed from multiple databases.RESULTS The expression level of SLC2A1 was up-regulated,but the expression level of SLC2A2 was down-regulated in HepG2 cells,HepG215 cells,and liver cancer patients.The expression levels of SLC2A1 and SLC2A2 were related to tumor volume,grade,and stage in HCC.Interestingly,the expression levels of SLC2A1 and SLC2A2 were negatively correlated.Further,high SLC2A1 expression and low SLC2A2 expression were linked to poor overall survival and relapse-free survival.SLC2A1,SLC2A2,and frequently altered neighbor genes played a major role in the occurrence and development of tumors.Notably,SLC2A1 was positively correlated with tumor immune infiltration,while SLC2A2 was negatively correlated with tumor immune infiltration.Particularly,SLC2A2 methylation was positively correlated with lymphocytes.CONCLUSION SLC2A1 and SLC2A2 are independent therapeutic targets for HCC,and they are quintessential marker molecules for predicting and regulating the number and status of immune cells in HCC.

Cathepsin L, transmembrane peptidase/serine subfamily member 2/4, and other host proteases in COVID-19 pathogenesis – with impact on gastrointestinal tract

Severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)seems to employ two routes of entrance to the host cell;via membrane fusion(with the cells expressing both angiotensin converting enzyme 2(ACE2)and transmembrane peptidase/serine subfamily member 2/4(TMPRSS2/4))or via receptor-mediated endocytosis(to the target cells expressing only ACE2).The second mode is associated with cysteine cathepsins(probably cathepsin L)involvement in the virus spike protein(S protein)proteolytic activation.Also furin might activate the virus S protein enabling it to enter cells.Gastrointestinal tract(GIT)involvement in SARS-CoV-2 infection is evident in a subset of coronavirus disease 2019(COVID-19)patients exhibiting GIT symptoms,such as diarrhea,and presenting viral-shedding in feces.Considering the abundance and co-localization of ACE2 and TMPRSS2 in the lower GIT(especially brush-border enterocytes),these two receptors seem to be mainly involved in SARS-CoV-2 invasion of the digestive tract.Additionally,in vitro studies have demonstrated the virions capability of infection and replication in the human epithelial cells lining GIT.However,also furin and cysteine cathepsins(cathepsin L)might participate in the activation of SARS-CoV-2 spike protein contributing to the virus invasiveness within GIT.Moreover,cathepsin L(due to its involvement in extracellular matrix components degradation and remodeling,the processes enhanced during SARS-CoV-2-induced inflammation)might be responsible for the dysregulation of absorption/digestion functions of GIT,thus adding to the observed in some COVID-19 patients symptoms such as diarrhea.

SCNH2 is a novel apelinergic family member acting as a potent mitogenic and chemotactic factor for both endothelial and epithelial cells

The gut hormone apelin is a major therapeutic focus for several diseases involving inflammation and aberrant cell growth. We investigated whether apelin-36 contained alternative bioactive peptides associated with normal physiology or disease. Amino acid sequence analysis of apelin-36 identified an amidation motif consistent with the formation of a secondary bioactive peptide (SCNH2). SCNH2 is proven to be mitogenic and chemotactic in normal/malignant cells and augments angiogenesis via a PTX-resistant/CT-X-sensitive G protein-coupled receptor (GPCR). Notably, SCNH2 is substantially more potent and sensitive than apelin-13 and vascular endothelial growth factor-A. Endogenous SCNH2 is highly expressed in human tumors and placenta and in mouse embryonic tissues. Our findings demonstrate that SCNH2 is a new apelinergic member with critical pluripotent roles in angiogenesis related diseases and embryogenesis via a non-APJ GPCR.