Effect of tanshinone IIA on the growth behavior of human hepatoma cell line BEL-7402 in vitro and its mechanism]

OBJECTIVE: To investigate the effect of tanshinone IIA on the growth behavior of human hepatoma cell line BEL-7402 in vitro and explore the mechanism. METHODS: Human hepatoma cell line BEL-7402 was exposed to tanshinoneIIA at different concentrations for 72 h, and the suppression of the cell growth was observed under inverted-phase contrast microscope. Apoptosis-related alterations in the cell morphology and biochemistry were examined under fluorescence microscope and transmission electron microscope (TEM) and by DNA agarose gel electrophoresis, and the apoptotic rate was quantified by flow cytometry (FCM). RESULTS: After treatment with 0-10 microg/ml tanshinone IIA for 72 h, the proliferation of BEL-7402 cells was significantly suppressed, and cell apoptosis occurred characterized by cell shrinkage, nuclear chromatin condensation and fragmentation, formation of membrane blebs and apoptotic bodies as observed under fluorescence microscope and TEM. DNA ladder was presented in DNA electrophoresis. FCM analysis yielded the cell apoptotic rates of (20.78+/-2.17) %, (24.64+/-2.07) %, (31.47+/-3.86) %, (43.65+/-4.04) % and (52.36+/-3.75) % at tanshinone IIA concentrations of 0.5, 1.0, 2.0, 5.0 and 10.0 microg/ml respectively, all significantly higher than those of the control group [(2.37+/-0.29)%]. CONCLUSION: Tanshinone IIA can inhibit the growth of human hepatoma BEL-7402 cells possibly through the mechanism of apoptosis induction.     

Growth Inhibition and Apoptosis Induction in Human Hepatoma Cells by Tanshinone Ⅱ_A

Summary:In order to study the effect of tanshinone Ⅱ_A on growth and apoptosis in human hepatomacell line BEL-7402 in vitro,the human hepatoma cell line BEL-7402 was treated with tanshinone Ⅱ_Aat various concentrations for 72 h.Growth suppression was evaluated by MTT assay;apoptosis-relat-ed alterations in morphology and biochemistry were ascertained under cytochemical staining(Hoechst33258),transmission electron microscopy(TEM),and DNA agarose gel electrophoresis.Apoptoticrate was quantified by flow cytometry(FCM).The results showed thst Tanshinone Ⅱ_A could inhibitthe growth of hepatoma cells in a dose-dependent manner,with IC_(50) value being 6.28μg/ml.Aftertreatment with 1—10 μg/ml tanshinone Ⅱ_A for 72 h,BEL-7402 cells apoptosis with nuclear chro-matin condensation and fragmentation as well as cell shrinkage and the formation of apoptotic bodieswere observed.DNA ladder could be demonstrated on DNA electrophoresis.FCM analysis showedhypodiploid peaks on histogram,and the apoptotic rates at 5     

Growth inhibition and apoptosis induction in human hepatoma cells by tanshinone II<Subscript>A</Subscript>

Summary In order to study the effect of tanshinone II_A on growth and apoptosis in human hepatoma cell line BEL-7402in vitro, the human hepatoma cell line BEL-7402 was treated with tanshione I_A at various concentrations for 72 h. Growth suppression was evaluated by MTT assay; apoptosis-related alterations in morphology and biochemistry were ascertained under cytochemical staining (Hoechst 33258), transmission electron microscopy (TEM), and DNA agarose gel electrophoresis. Apoptotic rate was quantified by flow cytometry (FCM). The results showed that Tanshinone II_A could inhibit the growth of hepatoma cells in a dose-dependent manner, with IC₅₀ value being 6.28 μg/ml. After treatment with 1–10 μg/ml tanshione II_A for 72 h, BEL-7402 cells apoptosis with nuclear chromatin condensation and fragmentation as well as cell shrinkage and the formation of apoptotic bodies were observed. DNA ladder could be demonstrated on DNA electrophoresis. FCM analysis showed hypodiploid peaks on histogram, and the apoptotic rates at 5 μg/ml concentration for 12 h, 24 h, 36 h, 48 h and 72 h were (2.32±0.16)%, (3.01±0.35)%, (3.87±0.43)%, (6.73±0.58)% and (20.85±1.74)% respectively, which were all significantly higher than those in the control group (1.07±0.13)%. It is concluded that Tanshione II_A could induce human hepatoma cell line BEL-7402 apoptosis, which may be related to the mechanism of growth inhibition. TANG Zhongzhi, male, born in 1966, Doctor in Charge This project was supported by a grant from Natural Sciences Foundation of Hubei Province (No. 2000J064).     

丹参酮ⅡA抑制HepG2细胞生长及诱导其凋亡的实验研究

目的:研究丹参酮ⅡA对人肝癌细胞HepG2的生长抑制作用和凋亡诱导作用.方法:以0μg/mL丹参酮ⅡA作阴性对照,MTT法检测0.5～10.0 μg/mL丹参酮ⅡA作用人肝癌细胞HepG2 24,48,72 h的生长抑制率;HT33258荧光染色、琼脂糖凝胶电泳、流式细胞仪检测不同浓度丹参酮ⅡA作用HepG2细胞72 h后的细胞凋亡.结果:0.5～10.0 μg/mL丹参酮ⅡA均能抑制人肝癌细胞HepG2生长,并有明显的时间和剂量依赖性;在24,48,72 h的半数抑制浓度分别为14.7,7.4,3.9 μg/mL;经丹参酮ⅡA作用后,荧光染色可以观察到典型的凋亡细胞形态特征;琼脂糖凝胶电泳结果显示除1.0 μg/mL组外均可见明显的凋亡细胞形成的梯状条带;流式细胞仪检测不同浓度丹参酮ⅡA作用72 h后的细胞凋亡率分别为20.32%±2.16%,28.01%±2.35%,33.87%±3.43%,46.73%±4.08%和57.85%±3.74%,与对照组比较差异均有统计学意义(P＜0.05).结论:丹参酮ⅡA在体外能明显抑制人肝癌细胞HepG2生长,抑制其生长的机制可能是诱导细胞凋亡.     

Antineoplastic mechanism of Octreotide actionin human hepatoma

Objectives To investigate whether apoptosis can be induced by Octreotide in human hepatoma cells in vitro and elucidate the antineoplastic mechanism of Octreotide in hepat oma. Methods A cultured human hepatoma cell line, BEL-7402, was exposed to Octreotide and ap optosis was evaluated by cytochemical staining (Hochesst 33 258), transmiss ion electron microscopy, agarose gel electrophoresis and flow cytometry (FCM).Results After exposure to 0.2 μg/ml Octreotide, apoptosis with nuclear chromatin cond ensation as well as fragmentation, cell shrinkage and the formation of apoptotic bodies was observed using cytochemical staining and transmission electron micros copy. A DNA ladder in agarose gel electrophoresis was also displayed. FCM show ed that the apoptotic cell number rose with an increase in the concentration of Octreotide (0-2 μg/ml). There was a positive correlation between Octreotide concentration and apoptotic rate in BEL-7402 cells (r=0.809, P&lt;0.05) .Conclusion Apoptosis in human hepatoma cells can be induced by Octreotide, which may be rel ated to the mechanism of antineoplastic action ofOctreotide in hepatoma.     

酒石酸锑钾在诱导人肝癌BEL-7402细胞凋亡中的影响

目的:本研究旨在明确酒石酸锑钾(PAT)在体外对人肝癌BEL7402细胞凋亡的影响及抑癌机制。方法:用PAT以不同浓度、不同时间作用于人肝癌BEL7402细胞,以诱导其凋亡。用MTT比色法观察其细胞毒性,荧光显微镜、透射电镜、TUNEL染色法及流式细胞术(FCM)等方法来检测凋亡,观察其形态学和生化方面的变化。结果:PAT以剂量依赖和时间依赖的方式抑制BEL7402细胞的生长。5～40μmol·L-1的PAT处理48h后,形态学上,肝癌细胞表现为细胞皱缩、核质浓缩、核碎裂、细胞起泡以及凋亡小体形式等凋亡特征的形态学改变。DNA末端原位标记染色法、流式细胞仪均能检测到凋亡细胞。结论:PAT在体外诱导肝癌BEL7402细胞凋亡,能作为一种凋亡诱导剂用于肝癌的治疗。     

亚砷酸体外对人肝癌细胞株BEL-7402影响的初步研究

目的 体外培养人肝癌细胞株BEL－7402,从多个角度探讨三氧化二砷（As2O3）的抗肿瘤作用及其机制。方法 应用倒置相差显微镜、电子显微镜、透谢电镜、流式细胞仪,分别对不同浓度加药组及对照组BEL－7402细胞的存活。形态学改变,细胞DNA含量的分布进行了观察和测定。结果 0.5、1、2μmol/L As2O3均能抑制人肝癌细胞株BEL－7402细胞的生长增殖。流式细胞仪分析显示,加药组在G1期细胞前均出现亚二倍体峰,且G0／G1期细胞减少,S期细胞增多;电镜下,对照组细胞核质比大、核大、核膜有明显切迹,0.5μmol/L As2O3组细胞核质比减少、核变圆、胞浆内出现分化良好的细胞器, 0.5、1、2μmol/L As2O3组均可见细胞膜完整、核固缩、凋亡小体形成。结论 三氧化二砷不仅抑制人肝癌细胞增殖,而且诱导细胞凋亡。     

去甲斑蝥素诱导肝癌细胞凋亡的实验观察

目的 观察去甲斑蝥素(NCTD)诱导肝癌细胞凋亡的分子学机制,为合理应用NCTD治疗肝细胞肝癌提供理论基础.方法 对肝癌细胞株SMMC-7721,BEL-7402进行体外培养,应用不同浓度的NCTD(3、10、30μg/ml)处理肝癌细胞,采用MTT方法检测细胞生存率,锥虫蓝拒染法检测细胞的死亡率,流式细胞仪检测细胞的凋亡率,Western印迹检测半胱氨酸天门冬氨酸蛋白酶(caspase)和聚腺苷二磷酸核糖聚合酶(PARP)蛋白,以及Bcl-2家族蛋白的表达.结果 MTT检测显示NCTD处理24、48和72 h后SMMC-7721细胞IC50分别为12、6和1.6μg/ml,BEL-7402细胞IC50分别为10、4和2 μg/ml;锥虫蓝拒染法结果显示,浓度为3、10、30 μg/ml的NCTD处理24 h后,SMMC-7721细胞的死细胞率分别为16.89%、46.16%、72.13%,均显著高于未处理对照组的死细胞率3.34%(P＜0.01);BEL-7402细胞的死细胞率分别为19.59%、50.10%、90.31%,均显著高于未处理对照组的死细胞率2.88%(P＜0.01).流式细胞仪检测显示,NCTD 10 μg/ml处理12 h,SMMC-7721细胞凋亡率达到27%,较未处理对照组(7%)升高20%,BEL-7402细胞凋亡率达到30%,较未处理对照组(8%)升高22%;Western印迹结果显示,NCTD 10μg/ml处理8、14和24 h后,在两株肝癌细胞均检测到caspase-9,-3活化,PARP蛋白裂解,以及Bcl-2、Bcl-XL和Mcl-1蛋白的表达下调.结论 结果提示NCTD具有较强的抑制肝癌细胞生存和诱导肝癌细胞凋亡作用.NCTD的抗肝癌作用机理可能是通过诱导多个Bcl-2抑凋亡家族蛋白表达下调,激活内源性线粒体凋亡通路,诱导肝癌细胞发生凋亡.     

绿脓杆菌制剂对人肝癌细胞株BEL-7402的杀伤效应

目的探索绿脓杆菌制剂对人肝癌细胞BEL-7402的杀伤效应。方法采用MTT法检测不同浓度的绿脓杆菌制剂对人肝癌细胞BEL-7402增殖的作用,同时利用电子显微镜、透射电镜观察细胞BEL-7402的形态学变化。结果MTT检测表明：绿脓杆菌制剂为10×107/mL、5×107/mL、2.5×107/mL时对肝癌细胞生长杀伤作用与对照组比较差异有高度统计学意义（P〈0.01）;电子显微镜、透射电镜观察发现肝癌细胞BEL-7402于12、24 h出现凋亡形态学改变,40 h形态学表现为凋亡与坏死并存。结论绿脓杆菌制剂对肝癌细胞BEL-7402生长有抑制作用,诱导细胞凋亡及坏死可能是主要作用机制。     

三氮唑席夫碱衍生物对肝癌细胞裂亡的影响

目的:研究3-吡啶-3-基-4-[(4-甲氧基-苯亚甲基)氨基]-5-甲硫基-1,2,4-三唑(LH-38)对肝癌细胞BEL-7402裂亡的作用。方法:BEL-7402细胞常规培养于RPMI-1640培养液中,细胞生长至对数生长期加LH-38(终浓度分别为1×10-4mol/L和1×10-5mol/L),连续培养48 h或72 h。四甲基偶氮唑蓝(MTT)比色法检测细胞增殖,荧光染料Hoechst33258和PI联染检测细胞死亡,免疫细胞化学法检测激活型Caspase-3表达。结果:LH-38抑制BEL-7402细胞增殖并呈浓度依赖关系,IC50为3.0×10-4mol/L;1×10-5mol/L浓度的LH-38处理细胞72 h,镜下可见多倍体细胞明显增多,可见微核或多核细胞染色体自发性的凝集,有丝分裂异常,存活或死亡的多核或单核巨细胞同时存在,存活细胞的激活型Caspase-3表达阴性;1×10-4mol/L浓度LH-38处理细胞48 h,可明显诱导细胞凋亡。结论:不同浓度LH-38可以引起人肝癌细胞BEL-7402细胞裂亡或细胞凋亡,即两种不同形式的细胞死亡。     

A study on apoptosis of nasopharyngeal carcinoma cell line induced by Tanshinone II A and its molecular mechanism]

OBJECTIVE: To assess effect of Tanshinone II A on the apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-1 and inquire into the mechanism there in involved. METHODS: The CNE-1 cells cultured in vitro were treated with 0.5 microgram/ml Tanshinone II A for 4 days. The morphology of the cells was observed by microscopy. The cell growth and proliferation were measured by cell counting. The DNA break was examined by gel electrophoresis. The cell cycle, apoptosis index (AI) and the expression of apoptosis associated gene were analysed by flow cytometry. RESULTS: 0.5 microgram/ml Tanshinone II A could induce the apoptosis of CNE-1 cells. In the treatment group, the morphologic characteristics of apoptotic cells were observed, the DNA of cells presented "ladder" break, the cell growth and proliferation were inhibited obviously, and the AI was 16.9% (the AI of control group was 6.4%). The cells were arrested in G0/G1 phase and cellular DNA synthesis was inhibited. The expressions of apoptosis associated gene fas, bax, p53 and p21 were up-regulated; the expression of bcl-2 was down-regulated. CONCLUSION: Tanshinone II A can induce apoptosis of human nasopharyngeal carcinoma cells. Its molecular mechanism may relate to modulation of the apoptosis associated gene expression.     

紫杉醇、5-氟脲嘧啶抑制肝癌细胞BEL-7402生长和诱导凋亡的比较

OBJECTIVE: To investigate the effect of paclitaxel and 5-flurouracil (5-Fu) on growth inhibition and apoptosis of human hepatoma BEL-7402 cells. METHODS: Growth inhibition of BEL-7402 cells treated with paclitaxel and 5-Fu, respectively, was measured by ATP-tumor chemosensitivity assay (ATP-TCA), and the cell cycle kinetics and apoptosis were analyzed by flow cytometry and microscopic examination. RESULTS: BEL-7402 cells were highly sensitive to paclitaxel with growth inhibition observed in both dose- and time-dependent manners (IC(50)=5.58 x 10(-7) mol/L). Paclitaxel induced significantly higher rate of cell apoptosis than the control group (P<0.05) but significantly lower rate than that induced by 5-Fu (P<0.01). Necrosis was observed predominantly in paclitaxel-treated cells whereas 5-Fu caused mainly cell apoptosis (P<0.05). Levels of apoptosis increased in proportion to the decrement of paclitaxel concentration but directly proportional to increment of 5-Fu concentration. CONCLUSIONS: Paclitaxel and 5-Fu are effective in inducing growth inhibition and apoptosis of BEL-7402 cells. While 5-Fu causes mainly apoptosis in hepatoma cells, the anticancer mechanism of paclitaxel is predominantly through induction of necrosis.     

家蝇抗菌肽Cecropin对人源性肿瘤细胞增殖与凋亡的影响

目的分析家蝇抗菌肽Cecropin对人源性肿瘤细胞体外生长增殖与凋亡的影响。方法采用四甲基偶氮噻唑蓝（MTT）MTT比色法测定家蝇抗菌肽Cecropin对人肺癌细胞株A549、人乳腺癌细胞株MCF-7、人宫颈癌细胞株Hela、人肝癌细胞株BEL-7402和人正常肝细胞株Changs,Liver生长增殖情况的影响,采用流式细胞术检测家蝇抗菌肽Cecropin作用后4株肿瘤细胞凋亡的情况,对照组不加家蝇抗菌肽Cecropin。结果家蝇抗菌肽Cecropin对4株人源性肿瘤细胞的生长均有抑制作用,并能诱导肿瘤细胞发生凋亡,但对人肝癌细胞BEL-7402作用效果相对较强。结论家蝇抗菌肽Cecropin能够影响人源性肿瘤细胞的生长与凋亡,作用机制需进一步研究。     

15-脱氧前列腺素J2体外诱导肝癌细胞失巢凋亡的研究

目的研究15-脱氧前列腺素J2（15-d-PGJ2）体外诱导人肝癌细胞系BEL-7402细胞失巢凋亡的特性,并探讨其机制。方法在纤连蛋白（Fn）或多聚2．羟乙基甲基丙烯酸脂（poly-HEMA）包被的培养板中,应用光学显微镜观察不同因素处理后的细胞生长状态和形态学变化;应用DNA片段分析和流式细胞术观察细胞的凋亡变化;应用Western印迹技术观察细胞焦点黏着斑激酶（FAK）和磷酸化FAK（p-FAK）蛋白质的表达;应用小RNA干扰技术来沉默FAK基因。结果在Fn包被的培养板中贴壁生长的BEL-7402细胞经15一d—PGJ：处理24h或48h后,细胞变圆,脱离细胞外基质呈悬浮状态改变,而且这种改变呈时间和剂量依赖性关系;其中以浓度为20μmol／L的15-d-PGJ2最为显著,24h和48h时问点细胞的黏附比率分别为（66．0±3．6）％和（35．0±5．0）％,与肝癌细胞对照相比差异有统计学意义（P〈0．05）。经流式细胞术和DNA片段分析后发现肝癌细胞发生了失巢凋亡;Western印迹显示p-FAK蛋白下降,而FAK蛋白质表达水平未发生改变。结论15-d-PGJ2在体外能够诱导BEL-7402细胞失巢凋亡,这一过程与细胞FAK磷酸化水平降低有关。     

紫杉醇、5-氟脲嘧啶抑制肝癌细胞BEL-7402生长和诱导凋亡的比较

目的 研究紫杉醇、5-氟脲嘧啶(5-Fu)抑制肝癌细胞BEL-7402增殖和诱导凋亡的效果。方法 采用ATP生物发光法检测肝癌细胞株的药物敏感性,以流式细胞术、形态学方法分析细胞凋亡和细胞周期。结果 BEL-7402对紫杉醇高度敏感,对5-Fu低度敏感,抑制作用均存在剂量、时间效应。紫杉醇组3种浓度诱导细胞凋亡率均明显高于无药对照组(P<0.05),但低于5-Fu组(P<0.01);紫杉醇组细胞死亡率显著高于5-Fu组(P<0.05)。紫杉醇诱导凋亡率随药物浓度降低而增加,而5-Fu诱导凋亡率随药物浓度降低而降低。结论 紫杉醇可抑制BEL-7402肝癌细胞增殖和诱导凋亡。5-Fu抑癌以诱导细胞凋亡为主,紫杉醇则以引起细胞坏死为主。     

野生型p53基因对人肺癌细胞SPC-A1凋亡的诱导

目的：观察外源性野生型p53基因特染对人肺腺癌细胞SPC-A1增殖和凋亡的影响。方法：用脂质体Lipofectamine将真核表达载体pCMV-p53和pCMV-p53mtl35分别转染SPC-A1细胞，MTT法检测转染前后的细胞生长情况，于转染后24、48、72h分别收集细胞，以流式细胞技术(FCM)检测细胞凋亡率，并进行染色体DNA断裂的测定。结泉：pCMV-p53转染后的SPC-A1细胞生长受到明显抑制，转染后24、48、72h的凋亡率分别为1．78％、24．11％、35．75％，转染后48、72h的细胞染色体断裂明显。结论：野生型p53基因瞬间转染可诱导肺癌细胞SPC-A1凋亡。     

丹参酮ⅡA对人乳腺癌细胞MDA-MB-231作用的实验研究

目的 研究丹参酮ⅡA(tanshinone ⅡA)对雌激素受体阴性人乳腺癌细胞(MDA-MB-231)的生长抑制作用及机制.方法 采用四甲基偶氮唑蓝(MTT法)、克隆形成试验检测丹参酮ⅡA对MDA-MB-231细胞的增殖抑制作用;倒置显微镜观察细胞形态;流式细胞术检测丹参酮ⅡA对MDA-MB-231细胞周期的影响.结果 丹参酮ⅡA处理后,肿瘤细胞增殖活性降低(P<0.05),半效抑制浓度(IC50)为0.125μg/ml,克隆形成率下降(P<0.05),均呈时间-剂量-效应关系;倒置显微镜观察到典型的细胞凋亡形态学特征性改变;细胞周期各相发生变化,G0/G1期细胞数增加(P<0.05),S期和G2/M期细胞数减少(P<0.05);并能诱导其凋亡.结论 丹参酮ⅡA使人乳腺癌细胞MDA-MB-231的增殖受到抑制,促进肿瘤细胞凋亡,具有抗肿瘤作用.     

姜黄素单体影响肝癌BEL-7402细胞凋亡及其周期蛋白D表达的研究

目的 采用细胞学实验观察3种姜黄素单体对肝癌细胞株BEL-7402的抑制增殖及诱导细胞凋亡的作用强度和机制.方法 以四甲基偶氮唑蓝(MTT)、Annexin Ⅴ-FITC双标记及流式细胞术(FCM)实验观察3种姜黄素对肝癌细胞株BEL-7402增殖的抑制作用与细胞周期变化,采用 Western blot实验分析3种姜黄素对肝癌细胞株BEL-7402细胞周期蛋白D表达的影响.结果 (1)3种姜黄素均可通过诱导肝癌细胞凋亡而有效抑制肝癌细胞生长增殖,存在时间与剂量效应,以姜黄素Ⅲ抑制效果最强.(2)3种姜黄素通过影响细胞周期生长的调控信号、降低细胞周期蛋白D表达量致使肝癌细胞停滞于G1/S期.结论 3种姜黄素均可通过抑制肝癌细胞周期蛋白D表达而诱导细胞凋亡,有效地抑制肝癌细胞生长增殖,具有很高的临床治疗价值.     

Effect of lidamycin on telomerase activity in human hepatoma BEL-7402 cells

Objective To investigate the effect of lidamycin (LDM) on telomerase activity in human hepatoma BEL-7402 cells under the condition of LDM inducing mitotic cell death and senescence. Methods Chromatin condensation was detected by co-staining with Hoechst 33342 and PI. Cell multinucleation was observed by Giemsa staining and genomic DNA was separated by agarose gel electrophoresis. Fluorescent intensity of Rho123 was determined for mitochondrial membrane potential. MTT assay and SA-β-gal staining were employed to analyze the senescence-like phenotype. The expression of proteins was analyzed by Western blot. Telomerase activity was assayed by telomerase PCR-ELISA. Results Mitotic cell death occurred in LDM-treated cells characterized by unique and atypical chromatin condensation, multinucleation and increased mitochondrial membrane potential. However, no apoptotic bodies or DNA ladders were found. In addition, apoptosis-related proteins remained nearly unaltered. Senescence-like phenotype was identified by increased and elongated size of cells, growth retardation, enhanced SA-β-gal activity and the changes of senescence-related protein expression. Telomerase activity markedly decreased (P<0.01) in LDM-treated hepatoma BEL-7402 cells. Conclusion Mitotic cell death and senescence could be triggered simultaneously or sequentially after exposure of hepatoma BEL-7402 cells to LDM. The decrease in telomerase activity may play a key role in the defective mitosis and aging morphology. Further investigation of detailed mechanism is needed.     

反义IGF-Ⅰ寡核苷酸转染对人肝癌细胞增生及凋亡的影响

为研究胰岛素样生长因子 I(IGF I)反义寡核苷酸转染对人肝癌细胞增生、分化及凋亡的影响 ,探讨寡核苷酸转染治疗肝癌的可行性 ,利用反义核酸技术 ,合成针对IGF I的寡核苷酸片段 ,利用脂质体包裹反义IGF I寡核苷酸片段瞬时转染人肝癌细胞系BEL 74 0 2细胞 ,MTT法检测细胞增生 ;放免法检测培养细胞上清中AFP、CEA的分泌量 ;采用末端标记 (Tunel)法检测细胞凋亡的变化。结果发现转染反义IGF I寡核苷酸可使人肝癌细胞系BEL 74 0 2细胞增生下降 ,AFP、CEA表达降低 ,凋亡细胞数量增多。反义IGF I寡核苷酸转染人肝癌细胞系BEL 74 0 2细胞可以降低细胞增生、减少细胞去分化并诱导肝癌细胞凋亡