DCE-MRI诊断原发性肝癌效能和TACE术后评估肿瘤活性应用价值研究*

目的 探讨动态对比增强磁共振成像(DCE-MRI)检查诊断原发性肝癌(PLC)患者的效能及评价疗效的价值。方法 2017年1月～2021年6月我院诊治的肝占位性病变患者68例,均接受DCE-MRI检查,并行穿刺细胞学检查诊断,应用临床流行病学基本技术评估MRI诊断的效能。所有PLC患者接受经皮肝动脉化疗栓塞术(TACE)治疗,术后再次接受MRI检查,评估疗效。结果 DCE-MRI检查显示PLC患者肝内特征性恶性肿瘤表现,T1WI序列为低信号,T2WI序列为高信号,动脉期明显强化,门脉期快速消退,偶见明显的肿瘤包膜;在68例肝占位性病变患者,经穿刺细胞学检查,诊断PLC患者64例,非肿瘤病变4例;DCE-MRI检查诊断的灵敏度、特异度、准确率、阳性和阴性预测值分别为95.2%、75.0%、89.7%、93.8%和25.0%;在TACE术后,MRI检查发现肿瘤被完全灭活18例(28.1%),部分灭活25例(39.1%),肿瘤稳定15例(23.4%),疾病进展6例(9.3%)。结论 DCE-MRI检查在诊断PLC和治疗后疗效判断方面有独特的作用,可指导临床制定治疗方案,价值很大。     

异丙酚持续静脉泵入麻醉对接受微波消融治疗的原发性肝癌患者血管内皮糖萼和脑代谢的影响

目的 探讨应用丙泊酚持续静脉泵注麻醉对接受手术治疗的原发性肝癌(PLC)患者血管内皮糖萼和脑代谢的影响。方法 2017年8月～2020年7月在我院行微波消融术治疗的PLC患者78例,其中一半患者接受异丙酚麻醉,另一半接受地佐辛麻醉。在术前(T0)、术后1 d(T1)和术后7 d(T2),行血气分析,计算脑氧摄取率(CEO₂),采用ELISA法检测硫酸乙酰肝素酶(HS)和多配体蛋白聚糖-1。结果 异丙酚麻醉组诱导时间和苏醒时间分别为(40.3±3.2) s和(6.5±0.8)min,显著短于地佐辛麻醉组【分别为(52.0±3.1) s和(9.8±1.1) min, P<0.05】;在T1和T2,异丙酚麻醉组HS分别为(17.3±2.1)pg/ml和(8.2±0.4)pg/ml,显著低于地佐辛麻醉组【分别为(37.2±5.6)pg/ml和(16.0±2.5)pg/ml,P<0.05】,血清多配体蛋白聚糖-1水平分别为(8.2±0.1)pg/ml和(3.8±0.1)pg/ml, 显著低于地佐辛麻醉组【分别为(11.7±1.4)pg/ml和(5.9±0.2) pg/ml,P<0.05】,CEO₂分别为(28.0±2.4)%和(32.7±3.7)%,显著高于地佐辛麻醉组【分别为(25.0±3.1)%和(29.7±4.1)%,P<0.05】。结论 应用异丙酚持续静脉泵注麻醉在接受微波消融治疗的PLC患者能抑制血管内皮糖萼的激活,改善脑代谢,提高麻醉效果,减少并发症的发生。     

Hsa-miR-4282对肝癌细胞系SMMC-7721生长的影响

目的  探讨Hsa-miR-4282在肝癌细胞系SMMC-7721中的表达及其对细胞生长的影响。方法 采用实时荧光定量PCR法检测Hsa-miR-4282在人正常肝上皮细胞系HL-7702和人肝癌细胞系MHCC97-H、SMMC-7721，以及 20例肝癌组织及其相应癌旁组织中的表达。采用瞬时转染法将Hsa-miR-4282 mimics（上调组）和Hsa-miR-4282 inhibitor（下调组）分别转染肝癌SMMC-7721细胞，上调组和下调组分别设置相应阴性对照组。转染后采用MTT法检测细胞增殖能力，平板克隆形成实验检测细胞克隆形成能力，流式细胞仪检测细胞凋亡能力。结果 Hsa-miR-4282在肝癌组织、肝癌细胞MHCC97-H及肝癌细胞SMMC-7721中的表达均低于癌旁组织及正常肝细胞HL-7702（P＜0.05）。MTT实验结果显示，Hsa-miR-4282上调后肝癌SMMC-7721细胞的OD值低于其阴性对照组，而下调后OD值高于其阴性对照组 （P＜0.05）。平板克隆形成实验显示，下调组的细胞克隆数高于其阴性对照组［（240±7） 个 vs （191±10） 个，P=0.005）］，而上调组细胞克隆数低于基阴性对照组［（146±10） 个 vs （193±12） 个，P=0.013）］。流式细胞仪检测结果显示，Hsa-miR-4282上调组细胞凋亡率较其阴性对照组升高［（23.89±1.89）% vs（16.6±1.14）%，P=0.009）］，下调组细胞凋亡率较期阴性对照组降低［（14.98±0.46）% vs （17.79±0.73）%，P=0.010］。结论 Hsa-miR-4282上调可抑制肝癌SMMC-7721细胞增殖，促进细胞凋亡，可能与肝癌的发病机制有关。     

PNPLA3对肝细胞和肝癌细胞脂质代谢的影响*

PNPLA3基因在人类和哺乳动物体内都有表达,并且在肝癌细胞中也有表达。PNPLA3在人体和小鼠体内突变后可导致肝细胞内甘油三酯的蓄积,肝细胞发生脂肪变性。在人类和大鼠肝癌细胞使突变型PNPLA3过表达,与过表达野生型PNPLA3细胞比,也会导致肝癌细胞脂质的聚集。本文将归纳总结以往关于PNPLA3基因对肝细胞和肝癌细胞脂质代谢的影响,并分别讨论了PNPLA3对人类肝细胞和肝癌细胞、大鼠和小鼠肝细胞及其肝癌细胞的影响。     

ALPPS治疗巨大原发性肝癌患者近远期疗效研究

目的 探讨联合肝脏分隔和门静脉结扎的二步肝切除术(ALPPS)治疗巨大原发性肝癌(PLC)患者的近远期疗效。方法 2016年1月～2018年2月我院诊治的98例巨大PLC患者,其中20例接受ALPPS治疗(A组),38例在经肝动脉化疗栓塞术( TACE)治疗后手术切除肿瘤(B组),和40例接受直接手术切除肿瘤(C组)。术后,行CT增强扫描,应用IQQA-Liver肝脏CT解读分析系统进行肝脏三维重建,测量全肝体积( TLV)、残余肝脏体积(RLV)、术后3个月肝脏体积(FRLV),计算肝增生体积和肝增生率。结果 术后,A组FRLV为(366.3±31.7)mL,显著大于B组【(185.2±16.4)mL或C组(181.9±14.2)mL,P＜0.05】,肝增生率为(68.8±6.4)%,显著高于B组【(21.4±2.0)%或C组(22.6±2.3)%,P＜0.05】; 术后15 d,A组血清白蛋白水平为(37.0±2.7)g/L,显著高于B组【(34.1±2.9)g/L或C组(34.0±3.2)g/L,P＜0.05】;A组2 a总生存率为70.0%,无进展生存率为55.0%,B组分别为55.3%和36.8%,C组分别为52.5%和42.5%,三组之间无显著性差异(P＞0.05)。结论 ALPPS可使巨大PLC患者术后剩余肝脏体积在短时间内迅速增加,加快了患者术后肝功能的恢复。     

蛇床子素诱导内质网应激抑制SMMC7721人肝癌细胞增殖的机制研究＊

目的 研究蛇床子素抑制肝癌细胞增殖的机制。方法 采用20 - 100 μM蛇床子素干预SMMC7721人肝癌细胞24 h、48 h和72 h，运用MTT方法检测细胞增殖；采用20 - 80 μM蛇床子素干预SMMC7721肝癌细胞24 h，运用流式细胞术检测细胞凋亡及Western blot检测Bax和Bcl-2蛋白表达；采用 60 μM蛇床子素干预SMMC7721肝癌细胞48 h，运用流式细胞术检测细胞周期，以实时荧光定量PCR技术检测GRP78、DDIT3、GADD34、ATF4、EIF2A、GDF15、CDKN1B、FOXO3、RICTOR、SGK1基因表达，Western blot方法检测CyclinD1、CyclinE1、p27^Kip1和CDK4细胞周期蛋白及GRP78、GRP94、PERK、XBP-1s、CHOP、p-eIF2α(Ser51)内质网应激蛋白表达。各实验均以0.1% DMSO为对照组，并以0.1 μM毒胡萝卜素为内质网应激诱导剂组。结果 与对照组比较，100 μM蛇床子素干预24 h显著抑制SMMC7721肝癌细胞增殖，其抑制率为36.76%（P < 0.05）；60 - 100 μM蛇床子素干预48 h后显著抑制SMMC7721肝癌细胞增殖，抑制率为30.84%-52.49%（P < 0.05）；40 - 100 μM蛇床子素干预72 h后显著抑制SMMC7721肝癌细胞增殖，抑制率为32.7%-73.15%（P < 0.05）。大于60 μM蛇床子素可诱导SMMC7721肝癌细胞晚期凋亡且显著抑制促凋亡蛋白Bax表达（P < 0.05）。60 μM蛇床子素干预SMMC7721肝癌细胞48 h后，G1期细胞数增加、G2期和S期减少，且CyclinD1、CyclinE1和p27^Kip1蛋白表达显著被抑制（P < 0.05）。20 - 80 μM蛇床子素处理SMMC7721肝癌细胞24 h后，均显著促进GRP78、DDIT3（CHOP）、ATF4、FOXO3、RICTOR和SGK1基因表达（P < 0.05）； 20 μM蛇床子素显著促进GADD34和EIF2A基因表达（P < 0.05）；80 μM蛇床子素显著促进GDF15基因表达（P < 0.05），且CHOP、XBP-1s蛋白表达显著增强。结论 蛇床子素通过诱导内质网应激反应以抑制SMMC7721肝癌细胞增殖，从而发挥抗肝癌活性。     

Inhibition of fatty acid synthase (FASN) affects the proliferation and apoptosis of HepG2 hepatoma carcinoma cells via the β-catenin/C-myc signaling pathway

Introduction and objectivesResearch in the last few years has proven that inhibition of fatty acid synthase (FASN) suppresses the migration and invasion of hepatoma carcinoma cells. This study aims to explore the effect of fatty acid synthase knockdown on the apoptosis and proliferation of HepG2 cells.Materials and methodsThe human liver cancer cell line HepG2 was cultured and then transfected with FASN-specific siRNA and negative control RNAi. After 48 h, cells and protein lysates were used for western blotting, CCK-8 (cell counting kit-8) assays, flow cytometry and other tests. To assess cell apoptosis, Bax, Bcl-2 and caspase-3 were detected; to assess proliferation, CDK4 (cyclin-dependent kinases 4) and P21 were detected; and to determine the signaling pathway involved, β-catenin and C-myc were also detected.ResultsInhibition of FASN in HepG2 cells can decrease proliferation and promote apoptosis. Flow cytometry and CCK-8 assays demonstrated that the apoptosis rate of FASN-specific siRNA-transfected cells was significantly increased compared to that of the control cells (p < 0.01). In addition, the cell cycle analysis revealed that FASN-specific siRNA-transfected cells induced G1 phase arrest (p < 0.05), but an increasing trend in G2 (p < 0.05).Compared with expression in negative RNAi-transfected cells, the expression of Bcl-2 and CDK-4 was reduced and the expression of Bax, caspase-3 and P21 was increased in FASN-specific siRNA-transfected cells (p < 0.05). Regarding the signaling pathway, the expression of β-catenin and C-myc was significantly reduced when compared to that in negative control cells (p < 0.05).ConclusionsInhibition of FASN suppressed the cell survival of HepG2 cells by inhibiting the β-catenin/C-myc pathway. This result suggests the potential treatment value of FASN for hepatoma carcinoma (HCC).     

α-干扰素和COX-2抑制剂对人肝癌细胞SMMC-7721细胞增殖的抑制作用研究

目的研究α-干扰素和环氧合酶(COX-2)抑制剂塞来昔布(Celecoxib)对人肝癌SMMC-7721细胞的生长抑制作用。方法采用MTT比色法观察不同浓度的塞来昔布和α-干扰素处理对肝癌细胞增殖的影响;通过荧光显微镜检测细胞凋亡。结果实验组与对照组相比,能抑制人肝癌SMMC-7721细胞的增殖,两种药物高浓度联合作用48 h后,对肝癌细胞的生长抑制率可达92.44%±0.98%。α-干扰素和塞来昔布对肝癌细胞的增殖抑制具有协同作用。荧光显微镜观察到细胞皱缩、核质浓缩、核碎裂以及凋亡小体形成等凋亡形态学改变。结论 COX-2抑制剂塞来昔布和α-干扰素对人肝细胞癌SMMC-7721细胞的增殖有协同抑制作用,同时能诱导其凋亡,并且呈剂量和时间依赖性,这提示α-干扰素和COX-2抑制剂塞来昔布对HCC的预防和治疗可能有积极意义。     

抗癌扶正方对人肝癌细胞SMMC-7721的Bcl-2基因表达的调控作用

目的:研究与分析抗癌扶正方对人肝癌细胞SMMC-7721的Bcl-2基因表达的调控作用。方法:首先培养人肝癌细胞SMMC-7721,然后使用噻唑蓝比色法(MTT比色法)检测抗癌扶正方对人肝癌细胞SMMC-7721增殖能力,使用流式细胞仪检测人肝癌细胞SMMC-7721抑制能力和Bcl-2基因表达水平。结果:通过检测发现,抗癌扶正方可以有效抑制人肝癌细胞SMMC-7721增殖。通过流式细胞仪检测发现,抗癌扶正方作用于人肝癌细胞SMMC-7721后,Bcl-2基因表达水平明显降低。结论:抗癌扶正方可以通过调控Bcl-2基因表达从而抑制人肝癌细胞SMMC-7721的活性。     

Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation

Aim:

Interferon-γ inducible protein 16 (IFI16), a DNA sensor for DNA double-strand break (DSB), is expressed in most human hepatocellular carcinoma cell (HCC) lines. In this study we investigated the re-localization of chromatin-bound IFI16 by Nutlin-3, a DNA damage agent, in HCC cells in vitro, and the potential mechanisms.

Methods:

Human HCC SMMC-7721 (wild-type TP53), Huh-7 (mutant TP53), Hep3B (null TP53) and normal fetal liver L02 cell lines were examined. DSB damage in HCC cells was detected via γH2AX expression and foci formation assay. The expression of IFI16 and IFNB mRNA was measured using RT-PCR, and subcellular localization and expression of the IFI16 protein were detected using chromatin fractionation, Western blot analysis, and fluorescence microscopy.

Results:

Treatment of SMMC-7721 cells with Nutlin-3 (10 μmol/L) or etoposide (40 μmol/L) induced significant DSB damage. In SMMC-7721 cells, Nutlin-3 significantly increased the expression levels of IFI16 and IFNB mRNA, and partially redistributed chromatin-bound IFI16 protein to the cytoplasm. These effects were blocked by pretreatment with pifithrin-α, a p53 inhibitor. Furthermore, Nutlin-3 did not induce ectopic expression of IFI16 protein in Huh-7 and Hep3B cells. Moreover, the association of IFI16 with chromatin and Nutlin-3-induced changes in localization were not detected in L02 cells.

Conclusion:

Nutlin-3 regulates the subcellular localization of IFI16 in HCC cells in vitro in a p53-dependent manner.