γδT细胞对肝癌细胞的杀伤作用

目的 探讨γδT细胞对肝癌细胞的杀伤作用.方法 异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)和rhIL-2活化正常人外周血单个核细胞(PBMC),用流式细胞仪分析活化后的γδT细胞含量,用ELISA法检测培养上清液中IFN-γ含量,用乳酸脱氢酶释放法测定活化γδT细胞的杀伤活性.结果 体外活化的PBMC在10 d时γδT细胞比例从扩增前(4.34±1.79)%增加到(55.65±6.88)%,PBMC经IPP刺激培养后IFN-γ分泌量较不含IPP的对照组有显著增高,至第7天达峰值,活化后的淋巴细胞对肝癌细胞有较高杀伤活性.结论 IPP作为刺激剂可获得大量γδT细胞,刺激PBMC后可分泌大量IFN-γ,γδT细胞在体外对肝癌细胞有较高的杀伤活性.     

双特异抗体CD3xAFP介导CIK细胞对AFP阳性肝癌细胞的杀伤应用

目的建立双特异抗体anti-CD3xanti-AFP介导CIK细胞对AFP阳性肝癌细胞的特异杀伤。方法利用anti-CD3抗体联合细胞因子IL-2活化人外周血淋巴细胞扩增CIK细胞,流式细胞仪分析细胞表型。柠檬酸三钠还原法制备金溶胶,物理吸附制备载荷anti-CD3和anti-AFP两种单抗的胶体金制剂,光镜检测抗体活性。AlarmaBlue法检测双特异抗体介导CIK细胞对肝癌细胞HepG-2和宫颈癌细胞Hela的杀伤活性。结果 14 d活化后的人外周血淋巴细胞鉴定为CIK细胞。抗体活性检测证实纳米金制剂保持了anti-CD3xanti-AFP的双特异抗体活性,在其介导下,CIK对特异靶细胞HepG-2杀伤效率为42%,提高11.4%(P<0.05),而对非特异靶细胞Hela杀伤无明显变化。结论金标双特异抗体anti-CD3xanti-AFP可以加强CIK细胞对AFP阳性肝癌细胞的杀伤作用。     

莲子心总生物碱对人肝癌细胞的抑制作用

摘 要 目的：研究莲子心总生物碱对人肝癌HepG2细胞的抑制作用。方法: 采用CCK-8试剂盒研究莲子心总生物碱对HepG2细胞生长的影响,运用流式细胞仪检测细胞的凋亡率。结果: 作用时间相同时,随着给药浓度的增加,莲子心总生物碱对HepG2细胞增殖的抑制率增加,呈现出剂量依赖性,72 h时,莲子心总生物碱对HepG2细胞的半抑制浓度（IC50）为1.501 μg·mL^-1;在同一浓度下,随着作用时间的延长,莲子心总生物碱对HepG2细胞增殖的抑制率呈上升趋势,其中72 h时,10 μg·mL^-1莲子心总生物碱HepG2细胞增殖的抑制率达到72%。不同浓度的莲子心总生物碱作用于人肝癌细胞后,HepG2细胞凋亡率明显升高,并呈剂量依赖性,与空白对照组比较,差异有统计学意义（P＜0.05）,其中20 μg·mL^-1莲子心总生物碱对HepG2细胞凋亡率达85.6%。结论:莲子心总生物碱能诱导细胞的凋亡,对人肝癌HepG2细胞的增殖有明显的抑制作用。     

酒精诱导肝癌细胞BEL-7402凋亡的作用研究

目的 体外观察不同浓度的酒精对肝癌细胞BEL-7402诱导凋亡的作用,为寻求肝癌的治疗提供新思路.方法 不同浓度的酒精与肝癌细胞BEL-7402通过体外培养,MTT法检测细胞增殖的变化;Hoechst 33258/PI 荧光双染色观察细胞形态学变化;蛋白质免疫印迹(Western blotting)半定量分析Caspase-3和p53和Bax的表达变化.结果 药物浓度在100～300 mmol/L的范围显著抑制肝癌细胞增殖,且抑制率随浓度和时间呈依耐性,Caspase-3,Bax和p53的蛋白表达量明显增加.药物浓度在100～300 mmol/L作用48 h,肝癌细胞皱缩,呈现凋亡各期改变,细胞凋亡率显著高于对照组(P<0.05).结论 不同浓度的酒精对肝癌细胞BEL-7402有诱导凋亡的作用,其抑制作用呈浓度和时间依赖性.     

昆布多糖硫酸酯对化疗药物治疗肝癌细胞的增敏作用

目的：探讨LAMS对肝癌细胞化疗的增敏作用。方法：以免疫组化法测定用LAMS前后肝癌细胞Bcl—2基因蛋白含量，以MTT法分别测定5—Fu、MTX、MMC、ADM、CTX与LAMS联合及单独治疗肝癌细胞的IC50以及有效作用时间。结果：LAMS使肝癌细胞Bcl—2基因蛋白表达下降，并使肝癌细胞对5—Fu、MTX、MMC、ADM、CTX的敏感性增加，有效作用时间延长。结论：LAMS可有效地增加肝癌细胞对5—Fu、MTX、MMC、ADM、CTX的敏感性。     

表阿霉素对肝癌细胞蛋白质表达影响的研究

目的 以体外培养的人体肝癌细胞为研究对象,运用蛋白质组学的研究方法,对比分析表阿霉素作用前后肝癌细胞细胞质蛋白质的差异表达情况,为抗癌药物作用机制的阐释提供信息.方法 体外培养的肝癌细胞(包括抗癌药物处理前后的肝癌细胞)以超离心分离细胞质,双向电泳分离细胞质蛋白质,图像分析药物处理前后肝癌细胞的差异表达蛋白质,MALDI-TOF-MS鉴定差异蛋白质.结果 从抗癌药物处理前后的肝癌细胞中筛选出5个差异表达蛋白质.这些差异蛋白质涉及到细胞的能量代谢、蛋白质合成、凋亡调控等方面.结论 抗癌药物作用后肝癌细胞细胞质中蛋白质的表达发生了许多改变,可为抗癌药物作用机制的阐释提供有价值的信息.     

RMP基因对肝癌细胞稳定性及迁移能力的影响

目的研究RMP基因对肝癌细胞稳定性及迁移能力的影响。方法将SMMC-7721细胞等量分为对照组和实验组。合成RMP基因的小干扰RNA真核表达载体,转染到实验组肝癌SMMC-7721细胞内,用G418筛选出实验组干扰RMP基因的细胞株,RT-PCR检测两组细胞内RMP基因的表达,划痕实验法检测两组细胞的迁移能力。结果成功构建稳定的RMP基因干扰细胞株,与未干扰细胞对照组比较,实验组细胞p21基因表达减弱,细胞迁移能力降低。结论 RMP基因有维持肝癌细胞基因组稳定的重要作用,对保持细胞的迁移能力也有作用。     

木霉菌醇对肝癌细胞CBRH7919凋亡及形态变化的影响

目的探讨木霉菌醇(trichomerol)对大鼠肝癌细胞CBRH7919的作用。方法采用SRB细胞活性检测法检测木霉菌醇对CBRH7919细胞活性的影响;用流式细胞仪检测CBRH7919对细胞周期和凋亡的影响;激光共聚焦显微镜观察吖啶橙荧光染色细胞形态变化。结果 SRB检测显示木霉菌醇对CBRH7919细胞作用具有浓度和时间依赖性;木霉菌醇作用后CBRH7919细胞周期S期比例上调,而G1期比例下调;凋亡率具有浓度依赖性;吖啶橙荧光染色发现,细胞有明显凋亡。结论木霉菌醇对肝癌细胞CBRH7919具有良好的抑制作用,并可诱导细胞凋亡。     

华蟾素增强细胞因子诱导的杀伤细胞对肝癌细胞的杀伤活性

目的观察华蟾素对细胞因子诱导的杀伤细胞(CIKC)的表型变化及其对肝癌的细胞毒作用。方法采集健康供者的外周血单个核细胞(MNC)培养,收集非贴壁细胞并加入白细胞介素2(IL-2)诱导培养CIKC,将华蟾素加入CIKC,用四甲基偶氮唑盐(MTT)法检测CIKC杀伤BEL7402肝癌细胞株的活性。结果CIKC经华蟾素加入培养后,CIKC细胞群的CD3+CD8+,CD3+CD56+细胞比例和杀伤活性较单纯的CIKC更高。结论华蟾素有助于CIKC的细胞毒活性。     

Octreotide inhibits proliferation and induces apoptosis of hepatocellular carcinoma cells

AIM: To study the effect of octreotide on cell proliferation and apoptosis in different hepatocellular carcinoma (HCC) cells and hepatocytes. METHODS: The proliferation of HCC cells (HepG2, SMMC-7721) and hepatocytes (L-02) was determined by MTT assay. Apoptosis was detected either by fluorescent staining, transmission electron microscopy or flow cytometry. The content of AFP in the supernatant of cultured HCC cells was determined by electrochemiluminescence immunoassay. The expression of SSTR subtypes was identified by RT-PCR. RESULTS: The proliferation of HCC cells and L-02 cells was inhibited significantly by octreotide (0.25, 0.5, 1.0, 2.0 and 4.0 mg/L). However, the apoptosis of HCC cells markedly increased in a concentration-dependent manner. Both the apoptosis index and the percentage of apoptotic cells in L-02 cells were significantly lower than those of HepG2 and SMMC-7721 cells. The content of AFP in the supernatant of cultured HepG2 cells treated with octreotide was also statistically reduced.     

Coronarin D induces apoptotic cell death through the JNK pathway in human hepatocellular carcinoma

Coronarin D, a diterpene derived from the rhizomes of Hedychium coronarium, has been used to treat inflammatory diseases. Coronarin D can exert strong anticancer effects through cell growth prevention and cell cycle arrest in many cancer cells. In this study, we investigated the molecular mechanism through which coronarin D suppresses cell proliferation and triggers cell death in human hepatocellular carcinoma (HCC) cells. Treatment of Huh7 and Sk‐hep‐1 cells with coronarin D resulted in a significantly increased loss of mitochondrial membrane potential, leading to the cleavage and activation of caspase‐9, caspase‐8, and caspase‐3 and changes in Bax, Bcl‐2, and Bcl‐xL protein levels. Coronarin D significantly induced autophagy by increasing the expression of Beclin‐1 and LC3‐II and reducing the expression of p62. Moreover, Huh7 and Sk‐hep‐1 cells exposed to coronarin D had decreased expression of phosphorylated AKT, p38, and ERK and increased expression of phosphorylated JNK. Exposure of cells to the JNK‐specific inhibitor SP600125 attenuated the apoptotic effects of coronarin D. Taken together, this is the first study to report that coronarin D may effectively inhibit cell growth through apoptosis. We have provided evidence indicating that coronarin D induces cell death through the upregulation of JNK mitogen‐activated protein kinases in human HCC cells.     

五味子甲素协同吉西他滨抑制肝癌细胞HepG2增殖

目的探究五味子甲素(deoxyschizandrin)联合吉西他滨(gemcitabin,GEM)对肝癌细胞HepG2增殖的影响及其可能的作用机制。方法CCK8法和克隆平板实验检测五味子甲素单药、GEM单药及联合用药对肝癌细胞HepG2增殖能力的影响;流式细胞术检测单药组及联合用药组中HepG2细胞的凋亡比例;Western blot法检测各组细胞中BCL2、BAX、pro-caspase3、cleaved-caspase3、pro-caspase9、cleaved-caspase9、β-catenin和TCF-4的表达。结果五味子甲素、GEM及联合用药对细胞HepG2的增殖均具有抑制作用,促进其凋亡,且联合用药组对HepG2细胞的作用明显强于单药组(P<0.05);Western blot结果表明,与对照组相比,五味子甲素、GEM和联合用药对pro-caspase3、pro-caspase9表达无明显影响,显著减少BCL2、β-catenin及TCF-4蛋白的表达(P<0.05),上调BAX、cleaved-caspase3、cleaved-caspase9蛋白的表达(P<0.05)。其中,联合用药比单药更为显著(P<0.05)。结论五味子甲素协同GEM抑制肝癌细胞HepG2中β-catenin/TCF-4通路,进而减少细胞增殖,诱导其凋亡。     

Nimbolide induced apoptosis by activating ERK‐mediated inhibition of c‐IAP1 expression in human hepatocellular carcinoma cells

Nimbolide is one of the major compounds from the leaves and flowers of the neem tree and exhibits antitumor properties on various cancer cells. However, no report has shown that nimbolide induces apoptosis in vitro and in vivo in human hepatocellular carcinoma cells. Our results indicated that it inhibited cell growth in Huh‐7 and PLC/PRF/5 cells. We also found that nimbolide induced cell death through the induction of G2/M phase arrest and mitochondrial dysfunction, accompanied by the increased expression of cleaved caspase‐7, caspase‐9, caspase‐3, caspase‐PARP, and Bax and decreased expression of Mcl‐1 and Bcl‐2. A human apoptosis antibody array analysis demonstrated that inhibition of the apoptosis family proteins (XIAP, c‐IAP1, and c‐IAP2) was one of the major targets of nimbolide. Additionally, nimbolide sustained activation of ERK expression. Moreover, pretreatment with U0126 (MEK inhibitor) markedly abolished nimbolide‐inhibited cell viability, induced cell apoptosis, ERK phosphorylation, cleaved caspase‐9, caspase‐3, cleaved‐PARP activation, and increased c‐IAP1 expression in Huh‐7 cells. An in vivo study showed that nimbolide significantly reduced Huh‐7 tumor growth and weight in a xenograft mouse model. This study indicated the antitumor potential of nimbolide in human hepatocellular carcinoma cells.     

复尔康注射液对人肝癌细胞株及裸鼠移植瘤的抑制作用

目的:研究复尔康注射液对人肝癌细胞株增殖及裸鼠移植瘤生长的影响。方法:采用CCK8试剂盒检测复尔康注射液对人肝癌细胞株SMMC-7721和BeL-7402增殖的抑制作用;建立SMMC-7721细胞裸鼠移植瘤模型,通过比较肿瘤体积(tumor volume,TV)、相对肿瘤体积(relative tumor volume,RTV)和相对肿瘤增殖率(T/C)考察复尔康注射液对肿瘤生长的影响。结果:复尔康注射液对SMMC-7721和BeL-7402的IC50分别为(61.51±1.58),(68.39±7.16)mg·L-1;不同剂量复尔康注射液能够不同程度地抑制裸鼠移植瘤的生长,与阴性对照组比较,P<0.05。低、中、高剂量复尔康注射液组(0.065,0.130,0.260 g·kg-1)T/C分别为57.75%,39.9%和44.08%。结论:复尔康注射液体外能抑制人肝癌细胞株的增殖,体内能明显抑制SMMC-7721细胞裸鼠移植瘤的生长。     

A kojic acid derivative promotes intrinsic apoptotic pathway of hepatocellular carcinoma cells without incurring drug resistance

Hepatocellular carcinoma is one of the most pervasive cancers with low prognosis, high frequency of recurrence, and metastasis. Studies conducted have focused on extricating novel strategies for successful treatment. Kojic acid and its derivatives are already proven to have depigmenting, anti‐inflammatory, and anti‐neoplastic properties. In the present study, kojic acid and its 10 distinct derivatives were tested on HEPG2 cell line for their possible anti‐cancer effect and seven of them were observed to be cytotoxic. Compound 6 was chosen to proceed as the IC₅₀ dosage for HEPG2 cells was lower in comparison with the other derivatives and kojic acid itself. Further experiments pointed out that intrinsic apoptotic pathway was triggered with the exposure of the cells to IC₅₀ concentration of the derivative as the treatment led to escalation of intracellular ROS, induction of TP53 gene, and activation of caspase 3/7. Pro‐apoptotic Jnk and Bax genes were not triggered suggesting that the apoptotic pathway advance through an alternative route. Complementary experiments are in need; howbeit, the current findings suggest that the derivative offers a novel promising approach against hepatocellular carcinoma as it is not detrimental to healthy cells within the concentrations applied, and it does not induce drug resistance.     

结节性肝癌超选择肝段栓塞临床疗效观察

目的:探讨应用节段性肝动脉化疗栓塞(STACE)治疗结节性肝癌的临床疗效。方法:经临床及影像诊断为结节性肝癌46例,共有瘤体58个,瘤体直径2.0~5.0 cm,平均直径4.2 cm,给予节段性肝动脉化疗栓塞。结果:1、2、3年累计复发率和生存率分别为16.25%、33.6%、43.5%和97.4%、79.5%、64.2%。结论:STACE治疗结节性肝癌可降低其并发症和复发率,肝功能不受损害或损害很轻,提高疗效及生存期,是治疗结节性肝癌安全有效的方法。