Efficacy of radiofrequency ablation combined with sorafenib for treating liver cancer complicated with portal hypertension and prognostic factors

BACKGROUND Patients with liver cancer complicated by portal hypertension present complex challenges in treatment.AIM To evaluate the efficacy of radiofrequency ablation in combination with sorafenib for improving liver function and its impact on the prognosis of patients with this condition.METHODS Data from 100 patients with liver cancer complicated with portal hypertension from May 2014 to March 2019 were analyzed and divided into a study group(n=50)and a control group(n=50)according to the treatment regimen.The research group received radiofrequency ablation(RFA)in combination with sorafenib,and the control group only received RFA.The short-term efficacy of both the research and control groups was observed.Liver function and portal hypertension were compared before and after treatment.Alpha-fetoprotein(AFP),glypican-3(GPC-3),and AFP-L3 levels were compared between the two groups prior to and after treatment.The occurrence of adverse reactions in both groups was observed.The 3-year survival rate was compared between the two groups.Basic data were compared between the survival and non-surviving groups.To identify the independent risk factors for poor prognosis in patients with liver cancer complicated by portal hypertension,multivariate logistic regression analysis was employed.RESULTS When comparing the two groups,the research group's total effective rate(82.00%)was significantly greater than that of the control group(56.00%;P<0.05).Following treatment,alanine aminotransferase and aspartate aminotransferase levels increased,and portal vein pressure decreased in both groups.The degree of improvement for every index was substantially greater in the research group than in the control group(P<0.05).Following treatment,the AFP,GPC-3,and AFP-L3 levels in both groups decreased,with the research group having significantly lower levels than the control group(P<0.05).The incidence of diarrhea,rash,nausea and vomiting,and fatigue in the research group was significantly greater than that in the control group(P<0.05).The 1-,2-,and 3-year survival rates of the research group(94.00%,84.00%,and 72.00%,respectively)were significantly greater than those of the control group(80.00%,64.00%,and 40.00%,respectively;P<0.05).Significant differences were observed between the survival group and the non-surviving group in terms of Child-Pugh grade,history of hepatitis,number of tumors,tumor size,use of sorafenib,stage of liver cancer,histological differentiation,history of splenectomy and other basic data(P<0.05).Logistic regression analysis demonstrated that high Child-Pugh grade,tumor size(6–10 cm),history of hepatitis,no use of sorafenib,liver cancer stage IIIC,and previous splenectomy were independent risk factors for poor prognosis in patients with liver cancer complicated with portal hypertension(P<0.05).CONCLUSION Patients suffering from liver cancer complicated by portal hypertension benefit from the combination of RFA and sorafenib therapy because it effectively restores liver function and increases survival rates.The prognosis of patients suffering from liver cancer complicated by portal hypertension is strongly associated with factors such as high Child-Pugh grade,tumor size(6-10 cm),history of hepatitis,lack of sorafenib use,liver cancer at stage IIIC,and prior splenectomy.

Silencing MFN2 Drives WNT/β-catenin Nucleation to Reduce Sorafenib Sensitivity in Hepatocellular Carcinoma Cells

Objective Mitofusin-2(MFN2)is a mitochondrial membrane protein that plays a critical role in regulating mitochondrial fusion and cellular metabolism.To further elucidate the impact of MFN2,this study aimed to investigate its significance on hepatocellular carcinoma(HCC)cell function and its potential role in mediating chemosensitivity.Methods This study investigated the effects of silencing and overexpressing MFN2 on the survival,proliferation,invasion and migration abilities,and sorafenib resistance of MHCC97-L HCC cells.Additional experiments were conducted using XAV939(aβ-catenin inhibitor)and HLY78(aβ-catenin activator)to further validate these findings.Results Silencing MFN2 significantly promoted the survival and proliferation of MHCC97-L cells,enhanced their invasion and migration capacities,increased the IC50 of sorafenib,reduced the percentage of TUNEL-positive cells,and decreased the expression of proapoptotic proteins.Additionally,silencing MFN2 markedly induced the nuclear translocation ofβ-catenin,increasedβ-catenin acetylation levels and enhanced the expression of the downstream regulatory proteins Snail1 and Vimentin while inhibiting E-cadherin expression.Conversely,overexpressing MFN2 reversed the effects observed in MHCC97-L cells mentioned above.The results confirmed that silencing MFN2 activated theβ-catenin/epithelial-mesenchymal transition(EMT)pathway and reduced the sensitivity of cells to sorafenib,which could be reversed by XAV939 treatment.Conversely,overexpression of MFN2 inhibited theβ-catenin/EMT pathway and increased the sensitivity of cells to sorafenib,which could be altered by HLY78.Conclusion Low expression of MFN2 in HCC cells promotes the nuclear translocation ofβ-catenin,thereby activating the EMT pathway and mediating resistance to sorafenib.

Tmem39b promotes tumor progression and sorafenib resistance by inhibiting ferroptosis in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)poses a significant threat to human health.Resistance to sorafenib in the chemotherapy of HCC is a common and significant issue that profoundly impacts clinical treatment.While several members of the transmembrane(TMEM)protein family have been implicated in the occurrence and progression of HCC,the association between TMEM39b and HCC remains unexplored.This study revealed a significant overexpression of TMEM39b in HCC,which correlated with a poor prognosis.Subsequent investigation revealed that RAS-selective lethal 3(RSL3)induced pronounced ferroptosis in HCC,and knocking down the expression of TMEM39b significantly decreased its severity.Similarly,following the induction of ferroptosis in HCC by sorafenib,knocking down the expression of TMEM39b also decreased the severity of ferroptosis,enhancing HCC tolerance to sorafenib.In conclusion,we propose that TMEM39b promotes tumor progression and resistance to sorafenib by inhibiting ferroptosis in HCC.

The flavonoids from Pentorum chinense Pursh.mediates ferroptosis to alleviate sorafenib-induced liver injury in BRL-3A cells

Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(PCP)has hepatoprotective properties.There are currently few reports on PCP’s protective impact and mechanism against sorafenib-induced liver injury.Methods:To create a liver injury model,sorafenib was administered to BRL-3A cells.Cell viability assays,immunofluorescence tests,Western blotting,real-time quantitative PCR,and high-content imaging systems were utilized to examine PCP’s effect and mechanism.Results:In this study,PCP treatment mitigated the liver damage caused by sorafenib by enhancing cell survival,lowering lipid reactive oxygen species and malondialdehyde levels,and elevating glutathione levels.In addition,PCP can enhance the protein expression of cystine/glutamate transporter xCT and glutathione peroxidase 4,reduce iron content and alleviate mitochondrial toxicity.Further mechanism studies revealed that PCP inhibited ferroptosis by promoting the production of nuclear factor E2-related factor 2 nuclear translocation and subsequently affecting target genes(HO-1 and NQO1).Conclusion:Together,PCP regulates the nuclear factor E2-related factor 2 pathway,which helps to lessen ferroptosis brought on by sorafenib.

Luteolin alleviates sorafenib-induced ferroptosis of BRL-3A cells through modulation of the Nrf2/GPX4 signaling pathway

Background:Luteolin is a flavonoid chemical that exists in a variety of medicinal and edible plants and holds many biologically active properties in liver protection,anti-cancer,antioxidants,anti-inflammatory,neuroprotective,etc.According to its hepatoprotective properties,luteolin was selected to co-treat with sorafenib,one of the approved protein kinase inhibitors,to reduce sorafenib-induced normal liver cell damage.Methods:The BRL-3A cell line was treated with sorafenib to establish a liver injury model,followed by luteolin treatment.The cell viability was detected,and the mechanism of action was detected by immunofluorescence,western blotting,and real-time quantitative PCR.Results:The research findings demonstrated that luteolin could increase cystine/glutamate transporter xCT(SLC7A11)and glutathione peroxidase 4(GPX4)expression and display a chelating effect on iron,which led to increased glutathione and decreased malondialdehyde,Fe^(2+) and lipid reactive oxygen species contents in BRL-3A cells,and the sorafenib-induced mitochondrial membrane potential decrease was also inhibited.In addition,when sorafenib caused the accumulation of lipid reactive oxygen species,luteolin could help release this oxidative stress by activating nuclear factor E2-related factor 2(Nrf2)and up-regulating the expression of the associated genes heme oxygenase 1(HO-1)and quinone oxidoreductase 1(NQO1).Conclusion:Therefore,luteolin may ameliorate sorafenib-induced ferroptosis by activating the Nrf2-associated pathway without any impact on sorafenib anti-cancer activity.It can be used as an adjuvant to sorafenib to reduce liver injury in patients with hepatocellular carcinoma.

Sorafenib plus transarterial chemoembolization vs sorafenib alone for patients with advanced hepatocellular carcinoma: A systematic review and meta-analysis

BACKGROUND Although the past decade has seen remarkable advances in treatment options for hepatocellular carcinoma(HCC),the dismal overall prognosis still envelops HCC patients.Several comparative trials have been conducted to study whether transarterial chemoembolization(TACE)could improve clinical outcomes in patients receiving sorafenib for advanced HCC;however,the findings have been inconsistent.AIM To study the potential synergies and safety of sorafenib plus TACE vs sorafenib alone for treating advanced HCC,by performing a systematic review and metaanalysis.METHODS This study was conducted following the PRISMA statement.A systematic literature search was conducted using the Cochrane Library,Embase,PubMed,and Web of Science databases.Data included in the present work were collected from patients diagnosed with advanced HCC receiving sorafenib plus TACE or sorafenib alone.Data synthesis and meta-analysis were conducted using Review Manager software.RESULTS The present study included 2780 patients from five comparative clinical trials(1 was randomized control trial and 4 were retrospective studies).It was found that patients receiving sorafenib plus TACE had better prognoses in terms of overall survival(OS),with a combined hazard ratio(HR)of 0.65[95%confidence interval(95%CI):0.46–0.93,P=0.02,n=2780].Consistently,progression free survival(PFS)and time to progression(TTP)differed significantly between the sorafenib plus TACE arm and sorafenib arm(PFS:HR=0.62,95%CI:0.40–0.96,P=0.03,n=443;TTP:HR=0.73,95%CI:0.64-0.83,P<0.00001,n=2451).Disease control rate(DCR)was also significantly increased by combination therapy(risk ratio=1.36,95%CI:1.02-1.81,P=0.04,n=641).Regarding safety,the incidence of any adverse event(AE)was increased due to the addition of TACE;however,no significant difference was found in grade≥3 AEs.CONCLUSION The combination of sorafenib with TACE has superior efficacy to sorafenib monotherapy,as evidenced by prolonged OS,PFS,and TTP,as well as increased DCR.Additional high-quality trials are essential to further validate the clinical benefit of this combination in the treatment of advanced HCC.

Charcoal Nanoparticles as a Delivery System for Doxorubicin and Sorafenib in Treatment of Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) is the most common type of liver cancer and one of the leading causes of cancer-related death worldwide. Advanced HCC displays strong resistance to chemotherapy, and traditional chemotherapy drugs do not achieve satisfactory therapeutic efficacy. The delivery of therapeutic compounds to the target site is a major challenge in the treatment of many diseases. Objective: This study aims to evaluate activated charcoal nanoparticles as a drug delivery system for anticancer agents (Sorafenib and Doxorubicin) in Hepatocellular Cancer Stem Cells. Method: The percent efficiency of entrapment (% EE) of the doxorubicin and sorafenib entrapped onto the activated charcoal was obtained by determining the free doxorubicin and sorafenib concentration in the supernatant-prepared solutions. Then the characterizations of nanoparticles were formed by determination of the particle size distribution, zeta potential, and polydispersity index (PDI). The anticancer activity of activated Charcoal, Doxorubicin-ACNP, sorafenib-ACNP, free doxorubicin, and free sorafenib solutions was measured based on cell viability percentage in HepG2 cell lines (ATCC-CCL 75). In vitro RBC’s toxicity of Doxorubicin/sorafenib loaded charcoal was estimated by hemolysis percentage. Results: The synthesized Doxorubicin-ACNP and Sorafenib-ACNP were evaluated and their physiochemical properties were also examined. Essentially, the percent Efficiency of Entrapment (EE %) was found to be 87.5% and 82.66% for Doxorubicin-ACNP and Sorafenib-ACNP, respectively. The loading capacity was 34.78% and 24.31% for Doxorubicin-ACNP and Sorafenib-ACNP. Using the Dynamic Light scattering [DLS] for the determination of the hydrodynamic size and surface zeta potential, a narrow sample size distribution was obtained of (18, 68, and 190 nm for charcoal, 105, 255, and 712 nm for doxorubicin, and 91, 295, and 955 nm for sorafenib), respectively. A surface charge of −13.2, −15.6 and −17 was obtained for charcoal, doxorubicin/charcoal, and sorafenib/charcoal nanoparticles. The cytotoxic activity of Doxorubicin-ACNP and Sorafenib-ACNP was evaluated in-vitro against HepG2 cell lines and it was observed that Drug loaded ACNP improved anticancer activity when compared to Doxorubicin or Sorafenib alone. Moreover, testing the toxicity potential of DOX-ACNP and Sorafenib-ACNP showed a significant reduction in the hemolysis of red blood cells when compared to Doxorubicin and Sorafenib alone. Conclusion: In conclusion, it is notable to state that this study is regarded as the first to investigate the use of Activated charcoal for the loading of Doxorubicin and Sorafenib for further use in the arena of hepatocellular carcinoma. Doxorubicin-ACNP and Sorafenib-ACNP showed noteworthy anticancer activity along with a reduced potential of RBCs hemolysis rendering it as an efficacious carrier with a low toxicity potential.

Sorafenib通过抑制TGF-β/Smad途径延缓肾纤维化的研究

目的探讨sorafenib减轻肾纤维化的作用及机制。方法用低、中、高剂量sorafenib分别给单侧输尿管梗阻(UUO)模型大鼠灌胃或干预经TGF-β1刺激的NRK-52E细胞。HE染色观察各组肾组织纤维化情况,免疫荧光染色检测肾组织及NRK-52E细胞α-SMA、E-cadherin的表达情况。用流式细胞术测定各组NRK-52E细胞周期。Western blot检测各组NRK-52E细胞中Smad3和p-Smad3的表达变化。结果 HE染色结果显示,与UUO模型组相比,sorafenib治疗组肾间质纤维化明显减轻,小管萎缩、炎细胞浸润较轻(P<0.05);与对照组比较,sorafenib治疗组E-cadherin在NRK-52E细胞和肾组织中表达均增加,而α-SMA表达均降低(P<0.05);流式细胞术分析发现细胞周期停滞于G0/G1期的细胞数明显增加,而进入G2、S期的细胞数明显减少(P<0.05);与对照组比较,sorafenib干预组p-Smad3蛋白在NRK-52E细胞中表达降低,且与sorafenib剂量呈正相关(P<0.05)。结论 sorafenib具有抗肾脏纤维化作用,主要通过TGF-β/Smad途径发挥作用,可能为治疗肾纤维化提供一种早期干预的新手段。

Sorafenib联合化疗治疗进展期肾癌Ⅱ期临床研究

[目的]探讨Sorafenib联合吉西他滨、5-Fu治疗晚期肾细胞癌的疗效及安全性。[方法]入组转移性肾细胞癌患者19例,其中既往细胞因子治疗失败者15例。化疗方案采用吉西他滨1g/m2,d1,8,5-Fu400mg/m2静脉推注d1,随后5-Fu2.1g/m246h化疗泵泵入,每4周为1个周期。同时Sorafenib治疗,400mg/次,口服,2次/d,持续使用至疾病进展或出现不可耐受的毒副反应。[结果]19例患者均可评价疗效。客观有效率37%(7/19),临床受益率79%(15/19)。6个月的PFS百分比为37%(7/19),并且该7例患者目前均无疾病进展。常见毒副作用为Ⅲ～Ⅳ度的骨髓抑制、Ⅱ度以上的手足综合征、Ⅱ度以上的胃肠道反应、Ⅱ度以上的皮疹、Ⅰ～Ⅱ度脱发、Ⅰ～Ⅱ度高血压。[结论]研究提示Sorafenib联合吉西他滨,5-Fu治疗晚期肾细胞癌表现出了较好的疗效和一定的安全性。

Sorafenib联合紫杉醇不同给药顺序作用于人肝癌细胞BEL-7402的效应

背景与目的:Sorafenib是一种多靶点的抗肿瘤药物,其治疗原发性肝癌的疗效已经初步得到证实。本研究应用Sorafenib与紫杉醇(paclitaxel,TAX)联合,观察两药不同的给药顺序作用于人肝癌细胞BEL-7402的不同效应,初步探讨产生这种差异的机制。方法:采用MTT法检测Sorafenib与紫杉醇单药作用于BEL-7402细胞的IC50。流式细胞术检测Sorafenib与紫杉醇不同给药顺序作用后,BEL-7402细胞周期和凋亡率的变化。Western blot检测不同给药顺序作用后的BEL-7402细胞Bcl-2的表达情况。结果:Sorafenib和紫杉醇单药作用于BEL-7402细胞48h的IC50分别为(2.43±0.32)μg/mL、(1.89±0.72)μg/mL。分析Sorafenib、紫杉醇单药,先用紫杉醇再加入Sorafenib,先用Sorafenib再加入紫杉醇及紫杉醇与Sorafenib同时作用后BEL-7402细胞周期及凋亡率的变化,结果显示:①Sorafenib主要将细胞阻滞在S期,而紫杉醇主要将细胞阻滞在G2-M期。②先用紫杉醇再给予Sorafenib组细胞S期及G2-M期均有延长,且较其他组获得较高的凋亡率(36.43±2.29)%(P<0.01)。Western blot显示先用紫杉醇再给予Sorafenib组BEL-7402细胞的Bcl-2表达的水平最低。结论:Sorafenib联合紫杉醇不同给药顺序作用于BEL-7402细胞,先应用紫杉醇诱导再序贯给予Sorafenib时细胞凋亡率更高;产生这种不同效应的可能机制为两种药物作用于不同的细胞周期,以及不同给药方法后的Bcl-2的表达水平不同。

Sorafenib对肝癌肝移植术后肿瘤转移复发大鼠的治疗作用

目的:探讨Sorafenib对肝癌肝移植术后肿瘤转移复发大鼠的治疗作用。方法:Walker-256癌细胞于Wistar大鼠门静脉注射,模拟大鼠肝癌肝移植术后肿瘤转移复发模型,术后随机分为4组(n=16):对照组、Sorafenib低剂量组、Sorafenib中剂量组、Sorafenib高剂量组,另选取5只正常Wistar大鼠作为正常组。Sorafenib各剂量组和对照组分别于术后20 d各随机选取8只,抽取静脉血,采用酶联免疫吸附方法(ELISA)检测大鼠血清中血管内皮生长因子(VEGF)含量,检测大鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、胆红素(TB)含量;取大鼠肝脏,常规甲醛固定、制作石蜡切片,HE染色,常规镜下观察;采用免疫组织化学方法计数肝脏微血管密度(MVD);观察Sorafenib各剂量组及对照组剩余8只大鼠生存时间。结果:Sorafenib各剂量组及对照组大鼠肝脏均出现了肿瘤转移复发,但Sorafenib各剂量组大鼠肝脏癌灶坏死程度较对照组明显,癌灶周边肝脏细胞受累坏死程度较对照组轻;Sorafenib各剂量组大鼠血清VEGF、ALT、AST、TB浓度均低于对照组(P<0.05);Sorafenib各剂量组大鼠肝脏MVD计数低于对照组(P<0.05);Sorafenib中、高剂量组大鼠存活率高于对照组(P<0.05)。结论:Sorafenib可以抑制肝癌肝移植术后肿瘤转移复发大鼠癌灶微血管生成,同时减轻癌灶对正常肝细胞的损害作用,延长肝癌肝移植术后肿瘤转移复发大鼠的生存时间,对肝癌肝移植术后肿瘤转移复发大鼠起到一定的治疗作用。

Sorafenib对前列腺癌DU145细胞的抑制作用的研究

目的观察索拉非尼(Sorafenib)对雄激素非依赖性前列腺癌DU145细胞的抑制作用。方法用不同浓度Sorafenib处理前列腺癌DU145细胞24、48和72h后,MTT法检测Sor-afenib对DU145细胞的抑制作用,流式细胞仪检测细胞凋亡变化,Western blot检测不同浓度Sor-afenib处理72h后DU145细胞内ERK和Bcl-2的表达。结果 Sorafenib能显著抑制DU145细胞的体外生长,呈时间与剂量依赖性。DU145细胞凋亡率随着Sorafenib剂量的增加而增大,具有良好的量效关系(P<0.01);Sorafenib处理DU145细胞72h后,ERK和Bcl-2蛋白的表达明显下调(P<0.01)。结论 Sorafenib抑制DU145细胞增殖、诱导细胞凋亡,可显著抑制雄激素非依赖性前列腺癌细胞的体外生长。

浅谈新型靶向药物Sorafenib对肿瘤治疗的研究

新型靶向药物Sorafenib是一种多靶点的生物制药,被临床前研究和临床实验证实具有广泛的抗肿瘤作用。本文从探讨了其作用机理,并总结了其在肾细胞癌,肝癌,非小细胞肺癌等癌症中的临床治疗及实验数据,认为其具有潜在的广谱抗肿瘤作用,为肿瘤的生物靶向治疗翻开了新的篇章。

新药sorafenib治疗晚期肾癌

20%～30%肾癌患者确诊时已出现转移,局限性肾癌患者术后也有20%～40%发生转移。传统化疗、放疗及糖皮质激素治疗均不能有效防止肾癌进展,转移性肾癌患者的预后较差,IL-2和IFN-α是目前仅有的免疫治疗方法,但疗效有限。近来,随着对肿瘤血管生成的有关机制的关注,针对以受体和调控血管生成信号分子为靶点的新药开发日渐成熟。首个口服多激酶抑制剂sorafenib,以丝氨酸-苏氨酸和受体酪氨酸激酶为作用靶点,抑制肿瘤细胞的增殖和肿瘤血管的形成。Ⅲ期临床研究显示,与安慰剂相比,本品可使患者无进展生存期延长1倍。本品于2005年12月经美国FDA快速审获批准用于晚期肾癌的治疗。

益气化瘀解毒方对MRP、GST-π和Topo Ⅱ基因在Sorafenib获得性耐药人肝癌QGY7702细胞表达的干预研究

目的探讨益气化瘀解毒方干预后对Sorafenib获得性耐药人肝癌QGY7702细胞(QGY7702/Sora)增殖及MRP、GST-π和Topo Ⅱ基因表达的影响。方法培养QGY7702/Sora细胞和QGY7702细胞,利用Cell Counting Kit-8(CCK-8)法检测Sorafenib对细胞的半数抑制率浓度(IC50值),计算耐药指数RI;观察益气化瘀解毒方对耐药细胞的增殖影响;采用荧光定量PCR检测药物干预前后2种细胞中MRP、GST-π和Topo Ⅱ基因表达水平。结果亲本细胞和耐药细胞Sorafenib的IC50值分别为(7.993±0.522)μmol/L和(19.651±1.216)μmol/L,RI约为2.5。益气化瘀解毒方可抑制耐药细胞的增殖活性。2种细胞的MRP、GST-π、Topo Ⅱ表达量无明显差异(P>0.05)。Sorafenib组可促进耐药细胞MRP 、GST-π基因的过表达(P<0.05),益气化瘀解毒方组可抑制GST-π基因的过表达(P<0.01),且联合Sorafenib可显著提高Topo Ⅱ基因的表达量(P<0.01)。结论 QGY7702/Sora细胞MRP、GST-π和Topo Ⅱ的表达水平与亲本细胞无显著差异。耐药细胞对Sorafenib敏感性降低与MRP、GST-π过表达相关,而益气化瘀解毒方拮抗Sorafenib耐药与抑制GST-π过表达相关。

Sorafenib联合吡喹酮对日本血吸虫病小鼠肝纤维化病理学的作用

目的观察Sorafenib在吡喹酮杀虫基础上对小鼠日本血吸虫病肝纤维化病理形态学的作用。方法 65只小鼠经腹壁感染日本血吸虫尾蚴,8周后形成血吸虫病肝纤维化模型,随机分为感染组(10只)、Sorafenib组(15只)、吡喹酮组(20只)和吡喹酮联合Sorafenib组(20只),另设正常对照组(5只)并分组给药。吡喹酮组:150 mg/(kg·d),连续治疗2天后给予等量生理盐水灌服2周;Sorafenib组:2天生理盐水灌服(剂量同吡喹酮组)后给予Sorafenib 2 mg/(kg·d),每日1次,灌服2周;吡喹酮联合Sorafenib组:吡喹酮150 mg/(kg·d),治疗2天后给予Sorafenib 2 mg/(kg·d),每日1次,灌服2周。正常组及感染对照组自给药开始每日1次灌服等量生理盐水至实验结束。处死小鼠后取肝脏,观察肝脏形态并取部分肝组织作病理学检查。应用HE染色观察小鼠肝脏的病理改变,Masson染色观察肝组织胶原纤维沉积范围,并采用IPP 6.0图文分析软件定量计算胶原纤维相对面积以反映纤维化程度。结果吡喹酮联合Sorafenib组小鼠肝脏病理损害明显减轻,纤维范围显著减少,该组胶原纤维相对面积(1.86±0.87)与感染组(5.97±1.77)和吡喹酮组(3.78±1.08)比较差异有统计学意义(P<0.0001),单用Sorafenib组胶原纤维相对面积(5.51±1.11)与感染组比较差异无统计学意义(P=0.42)。结论 Sorafenib在吡喹酮杀虫基础上能减轻日本血吸虫病肝脏病理损伤,二者联合有显著的增效作用。

Cryptotanshinone increases the sensitivity of liver cancer to sorafenib by inhibiting the STAT3/Snail/ epithelial mesenchymal transition pathway

Objective:Sorafenib resistance has been a major factor limiting its clinical use as a targeted drug in liver cancer.The present study aimed to investigate whether cryptotanshinone can enhance the sensitivity of liver cancer and reduce the resistance to sorafenib.Methods:Sorafenib-resistant cells were established based on HepG2 and Huh7 cell lines.And the anti-tumor effect of sorafenib combined with cryptotanshinone on the sorafenib-resistant cells was verified by MTT,colony formation,transwell assays and tumor growth xenograft model.Moreover,the effects of the combined treatment on the expression of phosphorylated(p)-STAT3,as well as epithelial mesenchymal transition(EMT)and apoptosis related proteins of cells were evaluated by western blot analysis.Results:It was identified that cryptotanshinone inhibited the viability of both HepG2 and Huh7 cells in a dose-and time-dependent manner,and decreased p-STAT3 expression rather than total STAT3 expression at a concentration of 40μmol/L.In the sorafenib-resistant cells,sorafenib in combination with cryptotanshinone markedly inhibited cell viability,invasion and migration compared with sorafenib alone.In contrast,increased p-STAT3 level by colivelin led to the inhibition of the synergistic effect of cryptotanshinone and sorafenib not only on cell viability,but also on EMT and apoptosis,suggesting that cryptotanshinone and sorafenib may act by downregulating STAT3 signaling.Further,the inhibition of carcinogenicity effect was also verified in xenografted tumor models.Conclusion:The present results indicated that cryptotanshinone could synergize with sorafenib to inhibit the proliferative,invasive,and migratory abilities of sorafenib-resistant cells by downregulating STAT3 signaling.

Sorafenib通过诱导铁死亡抑制DU145前列腺癌细胞增殖的分子机制

目的 研究Sorafenib通过诱导铁死亡来抑制DU145前列腺癌细胞增殖的机制。方法 分别用Sorafenib、铁死亡诱导剂RSL3和FIN56以及上述药物联合铁死亡抑制剂ferrostatin-1(Fer-1)处理前列腺癌细胞(DU145、PC3),采用MTT法检测细胞活力、流式细胞术检测细胞内脂质过氧化物含量的改变、Western blotting方法检测铁死亡标志蛋白SLC7A11和GPX4的表达水平。结果 Fer-1可以逆转Sorafenib对DU145前列腺癌细胞的增殖抑制作用,并且能够逆转Sorafenib处理导致的细胞内过氧化物含量增加;Sorafenib处理DU145和PC3细胞,细胞SLC7A11蛋白水平下调,但GPX4蛋白表达不变。结论 Sorafenib可作为铁死亡诱导剂抑制DU145细胞增殖,其机制可能是通过下调SLC7A11表达诱导细胞发生铁死亡。

Adjuvant sorafenib after heptectomy for Barcelona Clinic Liver Cancer-stage C hepatocellular carcinoma patients

AIM: To investigate the efficacy and safety of adjuvant sorafenib after curative resection for patients with Barcelona Clinic Liver Cancer(BCLC)-stage C hepatocellular carcinoma(HCC).METHODS: Thirty-four HCC patients, classified as BCLC-stage C, received adjuvant sorafenib for highrisk of tumor recurrence after curative hepatectomy at a tertiary care university hospital. The study group was compared with a case-matched control group of 68 patients who received curative hepatectomy for HCC during the study period in a 1:2 ratio.RESULTS: The tumor recurrence rate was markedly lower in the sorafenib group(15/34, 44.1%) than in the control group(51/68, 75%, P = 0.002). The median disease-free survival was 12 mo in the study group and 10 mo in the control group. Tumor number more than 3, macrovascular invasion, hilar lymph nodes metastasis, and treatment with sorafenib were significant factors of disease-free survival by univariate analysis. Tumor number more than 3 and treatment with sorafenib were significant risk factors of diseasefree survival by multivariate analysis in the Cox proportional hazards model. The disease-free survival and cumulative overall survival in the study group were significantly better than in the control group(P = 0.034 and 0.016, respectively). CONCLUSION: Our study verifies the potential benefit and safety of adjuvant sorafenib for both decreasing HCC recurrence and extending disease-free and overall survival rates for patients with BCLC-stage C HCC after curative resection.

放射联合Sorafenib对肝癌细胞的不同时效作用

【目的】在人肝癌细胞中观察放射联合Sorafenib对细胞生长的作用,并探讨其作用途径。【方法】选择人肝癌SMMC-7721和BEL-7402细胞株,分为单纯放射组(IR)、放射前30min联合Sorafenib组(IR+S-pre)和放射24h后联合Sorafenib组(IR+S-post)。细胞克隆形成实验检测照射后细胞克隆形成率,绘制细胞存活曲线并计算放射增敏比;细胞增殖抑制实验检测6Gy照射后第1、2、3、4、5、6d细胞增殖情况;免疫荧光染色观察照射后DNA损伤阳性细胞比例;流式细胞仪检测放射后细胞凋亡比例。【结果】细胞克隆形成实验结果显示,IR+S-pre放射后细胞克隆形成增加,放射增敏比在SMMC-7721和BEL-7402细胞中分别为0.78和0.88;而IR+S-post放射后细胞克隆形成减少,放射增敏比在SMMC-7721和BEL-7402细胞中分别为1.43和1.23。细胞增殖抑制实验显示IR+S-pre与IR放射后细胞增殖抑制率相似,而IR+S-post放射后细胞增殖抑制率明显提高。Sorafenib在SMMC-7721与BEL-7402中均有诱导细胞凋亡作用。IR+S-pre在放射后24h细胞凋亡比例明显增高[IRvsIR+S-pre:(6.1±1.0)%vs(18.3±2.0)%(SMMC-7721),(8.2±2.1)%vs(17.0±2.4)%(BEL-7402),P<0.001]。IR+S-post在放射后48h细胞凋亡比例明显增高,[IRvsIR+S-post分为(6.9±2.0)%vs(15.9±1.8)%(SMCC-7721),(8.0±1.5)%vs(14.2±2.5)%(BEL-7402),P<0.05]。放射后30min,IR+S-pre与IR的DNA受损阳性细胞比例无明显差异。放射后6hIR+S-pre的DNA受损阳性细胞残留比例明显下降[IRvsIR+S-pre:(59.9±2.4)%vs(23.8±2.9)%(SMMC-7721),(46.4±3.8)%vs(25.0±3.0)%(BEL-7402),P<0.001]。【结论】放射联合Sorafenib对肝癌细胞作用具有明显时效性,放射24h后联合Sorafenib增加放射引起的细胞克隆性生长抑制和增殖抑制,而放射前30min联合Sorafenib作用相反。