单抗F(ab′)2段导向抗肝癌阿霉素免疫毫微球的制备及其体外杀伤癌细胞作用

目的制备人肝癌特异性单克隆抗体HAb18的F(ab′)2片段导向抗肝癌阿霉素(ADR)白蛋白(HSA)免疫毫微球(HAb18 F(a b′)2-ADR-HSA-NP),并研究其体外靶向肝癌细胞和杀伤肝癌细胞的作用.方法采用乳化高温固化法制备阿霉素白蛋白毫微球(ADR-HSA-NP)后,应用改进的N-琥珀酰亚胺基-3-(2-吡啶二硫)丙酸酯(SPDP)法制备HAb18 F(ab′)2-ADR- HSA-NP.在光镜和电镜下观察体外HAb18 F(ab′)2-ADR-HSA-NP和ADR-HSA-NP与肝癌细胞株SMMC-7721的结合特性,并用3H-TdR法测定两种微球体外杀伤肝癌细胞株SMMC -7 721的作用.结果在荧光染色后,HAb18 F(ab′)2-ADR-HSA-NP表面发出明亮黄绿色荧光,而ADR-HSA-NP未见荧光.HAb18 F(ab′)2-ADR-HSA-NP能结合肝癌细胞株SMMC-7721,且能呈剂量依赖地有效杀伤肝癌细胞株SMMC-7721,而ADR-HSA-NP 不能结合和明显杀伤肝癌细胞株SMMC-7721.两种微球均不能结合和杀伤人大肠癌细胞株SW 1116.结论 HAb18 F(ab′)2-ADR-HSA-NP具良好的体外特异性结合并杀伤人肝癌细胞.     

单抗F（ab‘）2段导向抗肝癌阿霉素免疫毫微球的制备及其体外杀伤癌细胞作用

目的：制备人肝癌特异性单克隆抗体HAb18的F（ab‘)2片段导向抗肝癌阿霉素（ADR）白蛋白（HSA）免疫毫微球（HAb18F(ab‘)2-ADR-HSA-NP)，并研究其体外靶向肝癌细胞和杀伤肝癌细胞的作用。方法：采用乳化高温固化法制备阿霉素白蛋白毫微球（ADR－HSA－NP）后，应用改进的N－琥珀酰亚胺基－3－（2－吡啶二硫），丙酸酯（SPDP）法制备HAb18F(ab‘)2-ADR-HSA-NP。在光镜和电镜下观察体外HAb18 F(ab‘)2-ADR-HSA-NP和ADR－HSA－NP与肝癌细胞株SMMC－7721的结合特性，并用3H－TdR法测定两种微球体外杀伤肝癌细胞株SMMC－7721的作用。结果：在荧光染色后，HAb18F(ab‘2)-ADR-HSA-NP表面发出明亮黄绿色荧光，而ADR－HSA－NP未见荧洮，HAb18F(ab‘)2-ADR-HSA-NP能结合肝癌细胞株SMMC－7721，且能呈剂量依赖地有效杀伤肝癌细胞株SMMC－7721，而ADR－HSA－NP不能结合和明显杀伤肝癌细胞株SMMC－7721，两种微球均不能结合和杀伤人大肠癌细胞株SW1116。结论：HAb18F(ab‘)2-ADR-HSA-NP具良好的体外特异性结合并杀伤人肝癌细胞。     

抗人肝癌免疫毫微粒的制备及体外免疫学性质的鉴定

目的研究抗人肝癌免疫毫微粒的制备以及对靶细胞的杀伤作用。方法使用异型双工能SPDP对人肝癌以及载阿霉素（ADR）和人血清蛋白微粒进行偶联,通过实验对其的免疫特性进行测试。检测的方法使用MTT法,这种方法的检测对免疫毫微粒是否能够对体外具备杀伤的作用十分有效。检测后在人肝癌老鼠模型上使用HAb18-HSA（ADM）-NS、HSA（ADM）-NS和ADM,来测试其对肝癌的抑制情况。结果 HAb18抗体可以偶联到毫微粒上,对体外杀伤的细胞值为45.5μg/mL,与ADM的366.1μg/mL比较下,明显了降低。免疫毫微粒能够和靶细胞结合并且对靶细胞具有杀伤和选择功能。结论偶联的方法适用制备肝癌免疫毫微粒,并且免疫毫微粒在对抗肝癌治疗上有着明显的效果,值得推广使用。     

单抗F(ab′)2段导向抗肝癌阿霉素免疫毫微球的制备及其体外杀伤癌细胞作用

目的　制备人肝癌特异性单克隆抗体 HAb18的 F(ab′) 2 片段导向抗肝癌阿霉素 (ADR)白蛋白(HSA)免疫毫微球 (HAb18F(ab′) 2 - ADR- HSA- NP) ,并研究其体外靶向肝癌细胞和杀伤肝癌细胞的作用。方法　采用乳化高温固化法制备阿霉素白蛋白毫微球 (ADR- HSA- NP)后 ,应用改进的 N-琥珀酰亚胺基 - 3- (2 -吡啶二硫 )丙酸酯 (SPDP)法制备 HAb18F(ab′) 2 - ADR- HSA- NP。在光镜和电镜下观察体外 HAb18F(ab′) 2 - ADR- HSA- NP和 ADR- HSA- NP与肝癌细胞株 SMMC- 772 1的结合特性 ,并用 3H- Td R法测定两种微球体外杀伤肝癌细胞株SMMC- 772 1的作用。结果　在荧光染色后 ,HAb18F(ab′) 2 - ADR- HSA - NP表面发出明亮黄绿色荧光 ,而 ADR-HSA- NP未见荧光。 HAb18F(ab′) 2 - ADR- HSA- NP能结合肝癌细胞株 SMMC- 772 1,且能呈剂量依赖地有效杀伤肝癌细胞株 SMMC- 772 1,而 ADR- HSA- NP不能结合和明显杀伤肝癌细胞株 SMMC- 772 1。两种微球均不能结合和杀伤人大肠癌细胞株 SW1116。结论　 HAb18F(ab′) 2 - ADR- HSA - NP具良好的体外特异性结合并杀伤人肝癌细胞     

免疫毫微粒的内化作用及其逆转人肝癌细胞多药耐药性的研究

目的：观察免疫毫微粒能否内化进入人肝癌细胞及其多药耐药性(MDR)细胞，研究免疫毫微粒能否逆转MDR。方法：将抗人肝癌阿霉素白蛋白免疫毫微粒(HAbl8-ADR-HSA-NP)与人肝癌株SMMC-7721细胞及其MDR细胞(SMMC-7721／MDR^ )共同孵育，采用扫描电镜，激光共聚焦显微镜，透射电镜技术观察免疫毫微粒与靶细胞的结合及免疫毫微粒的内化过程；采用MTT比色分析法测定免疫毫微粒对靶细胞的杀伤率。计算免疫毫微粒对靶细胞的IC50和MDR细胞的耐药倍数(RF)。结果：激光共聚焦显微镜观察，SMMC-7721细胞表面和胞浆中均有许多荧光毫微粒；透射电镜观察。免疫毫微粒与SMMC-7721细胞及其MDR细胞在37℃孵育后，胞浆中可见有免疫毫微粒，随时间的延长细胞变性损伤越严重；当SMMC-7721及其MDR细胞预先经HAbl8抗体处理，再与HAbl8-ADR-HSA-NP孵育，则胞浆中未见免疫毫微粒；扫描电镜显示，多个HAbl8-ADR-HSA-NP紧密结合在SMMC-7721／MDR^ 表面。MDR细胞对于免疫毫微粒的耐药倍数(2.1)较其对于游离ADR(4．4)明显降低。结论：人肝癌特异性阿霉素白蛋白免疫毫微粒通过抗体介导能特异性内化进入人肝癌SMMC-7721细胞及其MDR细胞；该免疫毫微粒能增强MDR细胞对阿霉素杀伤的敏感性。有一定程度逆转MDR作用。     

阿霉素人血清白蛋白微球的制备及其性质的初步研究

目的 制备载阿霉索人血清白蛋白微球(ADR-HSA-MS)，并研究其药剂学性质和体外对肿瘤细胞的细胞毒作用。方法 以阿霉索(ADR)、人血清白蛋白(HSA)为材料，采用乳化高温固化法制备ADR-HSA-MS；采用光镜、电镜技术观察ADR-HSA-MS的形态；采用胃蛋白酶消化法、动态透析法分别测定ADR-HSA-MS的载药量，体外释药性质；采用MTT比色分析法测定ADR-HSA-MS对人肝癌株SMMC-7721细胞的细胞毒作用。结 ADR-HSA-MS圆球状，表面较光滑，平均粒径为1．2μm，外观为红色；ADR-HSA-MS表观载药量为2．73％，有效载药量为1．69％，释药呈持续缓慢状态，至5d时，其累积释药分数达50％，最大累积释药分数为65％；ADR-HSA-MS与SMMC-7721人肝癌细胞孵育4d后，对SMMC-7721细胞具明显杀伤作用，在0．3～2．5μg／ml ADR质量浓度范围内呈一定剂量依赖性，其杀伤曲线与游离ADR接近。结论 采用乳化高温固化法能制备出具缓释性质的载阿霉素人血清白蛋白微球。     

阿霉素免疫毫微粒对多药耐药肝癌的体内杀伤作用

目的 探讨阿霉素免疫毫微粒对多药耐药(MDR)肝癌的体内杀伤作用及其可能的机制.方法 用人肝癌细胞株耐阿霉素亚株(SMMC-7721/ADM)制成肝癌裸鼠模型,分别使用阿霉素免疫毫微粒、阿霉素毫微粒及阿霉素,检测三者的肿瘤抑制率:同时检测三者在肿瘤组织中的药物浓度.结果 阿霉素免疫毫微粒有比普通阿霉素毫微粒及阿霉素更强的体内逆转MDR的功能:阿霉素免疫毫微粒比阿霉素毫微粒及阿霉素在肿瘤中的药物浓度明显高.结论 免疫毫微粒有体内逆转MDR的功能,其可能机制是免疫毫微粒在体内可与肿瘤细胞紧密结合,所载药物释放后形成肿瘤组织内有较高的药物浓度.

Abstract：

Objective To observe the effect of the immunonanoparticles loaded with adriamycin in reversing multidrug resistance (MDR) in liver cancer in a nude mouse model and explore the possible mechanisms. Methods The cytotoxicity of adriamycin, adriamycin-loaded nanoparticles, and adriamycin-loaded immunonanoparticles was assessed in a nude mouse model bearing implant tumors of adriamycin-resistant hepatoma cell line SMMC-7721/ADM. The concentration of adriamycin in the tumor tissue was determined. Results Adriamycin-loaded immunonanoparticles showed significantly stronger cytotoxicity against the implant tumors of SMMC-7721/ADM than adriamycin-loaded nanoparticles and adriamycin.Administration of adriamycin-loaded immunonanoparticles resulted in significantly higher drug concentrations in the tumor tissue than adriamycin-loaded nanoparticles and adriamycin. Conclusion Adriamycin-loaded immunonanoparticles may reverse the MDR of liver cancers in vivo probably resulting from the close binding of the particles with the tumor cells to produce a high local concentration of adriamycin in the tumors.     

米托蒽醌聚氰基丙烯酸正丁酯毫微球抗肝癌效果的初步观察

作者用常位(肝内)及异位(右腋皮下)移植人肝癌动物瘤株(LTNM_4)裸小鼠模型,以阿霉素(ADR)、米托蒽醌(DHAQ)作比较,对肝靶向制剂米托蒽醌毫微球(DHAQ-PBCA-NS)的抗肝癌效果进行了研究。结果表明:ADR、DHAQ和DHAQ-PBCA-NS对常位移植人肝癌的抑瘤率分别为60.07％、67.49％和99.44％。对异位移植人肝癌的抑瘤率分别为80.03％、86.18％和92.90％。用抗增殖细胞核抗原(PCNA)单克隆抗体PC_(10),采用ABPAP法显示细胞增殖活性,ADR、DHAQ和DHAQ-PBCA-NS的PCNA计数分别为85.5±12.7％、76.0±13.4％和31.0±23.0％。经t检验显示,DHAQ抗肝癌效果与ADR相当,而DHAQ-PBCA-NS抗常位移植人肝癌的效果明显优于ADR和DHAQ,具有应用研究前景。     

柴胡注射液对人肝癌细胞株SMMC-7721增殖及凋亡的影响

目的:探讨柴胡注射液对人肝癌细胞株SMMC-7721凋亡的影响及相关机制。方法:将人肝癌SMMC-7721细胞株(由西安交通大学医学院第一附属医院临床分子试验中心提供)分为空白对照组(加10%小牛血清的1640培养液)、氟尿嘧啶阳性对照组(10μg/ml)及实验组(分别为5mg/ml、10mg/ml、20mg/ml、25mg/ml、50mg/ml柴胡注射液)。采用噻唑蓝比色法(即MTT)检测人肝癌细胞株SMMC-7721对柴胡注射液药物和氟尿嘧啶注射液的敏感性;应用流式细胞术(FCM)检测柴胡注射液对人肝癌细胞株(SMMC-7721细胞)周期的影响。结果:柴胡注射液对SMMC-7721的生长有明显的抑制作用,抑制率与柴胡注射液浓度呈正相关。不同浓度柴胡注射液处理肝癌细胞时,药物浓度与凋亡细胞且呈剂量依赖关系。结论:柴胡提取物可抑制人肝癌SMMC-7721细胞增殖,诱导其凋亡,作用机制可能与调控细胞周期有关。     

抗人大肠癌免疫磁性白蛋白微球的抗癌效果

目的:观察自制丝裂霉素C免疫磁性白蛋白微球(MMC-IMAMS)体外及体内抗大肠癌效果。方法:将丝裂霉素C免疫磁性白蛋白微球(MMC-IMAMS)与丝裂霉素C磁性白蛋白微球(MMC-MAMS)、单纯丝裂霉素C(MMC)比较,MTT法分别检测其体外对大肠癌细胞株LoVo的杀伤作用,然后通过移植人大肠癌裸鼠模型检测体内的肿瘤抑制率。结果:体外实验丝裂霉素免疫磁性白蛋白微球较单纯丝裂霉素磁性白蛋白微球有更强的肿瘤细胞抑制效率;裸鼠模型体内实验丝裂霉素免疫磁性白蛋白微球组肿瘤抑制率较丝裂霉素磁性白蛋白微球组和单纯丝裂霉素组具有显著差异。且丝裂霉素免疫磁性白蛋白微球是安全的,在体外磁场作用下可较长时间内沉积于靶区。结论:丝裂霉素免疫磁性白蛋白微球安全性好,靶向性更强大,体内体外均有更强的抗癌效果。     

Preparation and in vitro killing effect of adriamycin-loaded immunonanosphere against hepatoma led by F (ab')2 Fragment of monoclonal antibodies]

OBJECTIVE: To study the preparation method and in vitro killing effect of adriamycin (ADR)-loaded human serum albumin (HSA) immunonanosphere (HAb18 F(ab')2-ADR-HSA-NP) against hepatoma led by F(ab')2 fragment of human hepatoma specific monoclonal antibody HAb18. METHODS: After ADR loaded HSA nanosphere (ADR-HSA-NP) was prepared in the emulsifying high temperature solidifying way, HAb18 F(ab')2-ADR-HSA-NP was prepared using the modified N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) method. In vitro binding characters of HAb18 F(ab')2-ADR-HSA-NP and ADR-HSA-NP and hepatoma cell SMMC-7721 were observed under optical microscopy and electronic microscopy. In vitro effects of killing hepatoma cell SMMC-7721 of two microspheres were determined using the method of 3H-TdR. RESULTS: The surfaces of HAb18 F(ab')2-ADR-HSA-NP gave out bright yellow-green fluorescence after it was dyed with fluorescent agent, whereas ADR-HSA-NP did not give out fluorescence. HAb18 F(ab')2-ADR-HSA-NP could integrate with hepatoma cell SMMC-7721 and effectively killed hepatoma cell SMMC-7721 with dose dependence, but ADR-HSA-NP could not obviously integrate and kill SMMC-7721. Neither of the two microspheres could bind and kill human large intestine cancer cell SW1116. CONCLUSION: HAb18 F(ab')2-ADR-HSA-NP has a good character for in vitro specific targeting to bind and kill human hepatoma cell.     

survivin反义寡核苷酸对肝癌耐药细胞的作用及与化疗的关系

目的研究survivin反义寡核苷酸对人肝癌耐药细胞株的增殖和凋亡情况,以及对阿霉素化疗敏感性的影响。方法将人肝癌耐药细胞系SMMC-7721/ADM分为脂质体转染组、阿霉素组、正义寡核苷酸转染组、正义寡核苷酸转染 阿霉素组、反义寡核苷酸转染组、反义寡核苷酸转染 阿霉素组共6组。MTT法检测细胞相对存活率,流式细胞仪分析细胞凋亡率变化,RT-PCR和Westernblot印迹法检测细胞survivinmRNA和蛋白表达。结果survivin-ASODN作用后的人肝癌耐药细胞SMMC-7721/ADM的细胞凋亡率明显增高(P<0.05)。survivinmRNA和survivin蛋白表达:脂质体转染对照组、阿霉素组、正义寡核苷酸转染对照组、正义寡核苷酸转染对照 阿霉素组之间survivin蛋白表达无明显差异(P>0.05)。400ng/mlsurvivingASODN组和400ng/mlASODN 阿霉素组survivinmRNA较其他4组显著降低(P<0.05),但其两组之间表达无明显差异(P>0.05)。结论survivin反义寡核苷酸能降低肝癌耐药细胞survivin表达,增强人肝癌耐药细胞对阿霉素的化疗敏感性。     

猫人参注射液体外抗肝癌实验研究

目的:观察单味猫人参注射液在体外对肝癌细胞的抑制作用.方法:噻唑蓝还原法(MTT法)检测猫人参注射液对体外培养的人肝癌细胞株SMMC-7721、小鼠肝癌细胞株H22、大鼠肝癌细胞株CBRH-7919的生长抑制作用.结果:体外实验结果表明:250mg/ml浓度猫人参注射液对H22、CBRH-7919、SMMC-7721肝癌细胞株均有抑制作用,抑制率分别63.68%、41.51%、32.92%,IC50分别为:107.70mg/ml、519.49mg/ml、667.56mg/ml;在24h,48h,72h三个时相,对小鼠H22肝癌细胞的生长抑制作用存在较明显的时效关系,在72h内16.88mg/ml～250mg/ml浓度内药物浓度与抑制作用有正相关趋势,其相关系数为0.91.结论:猫人参注射液对不同肝癌细胞株有一定的抑制作用.     

Internalization of monoclonal antibodies against human hepatoma in the target cells]

The monoclonal antibodies against human hepatoma, HAb23, HAb18 and HAb8, were linked with 5 nm colloid gold. They were used to incubate with the target cells, QGY-7703 and SMMC-7721 human hepatoma cell lines, at 4 degree C for 1 hour. The incubated hepatoma cells were divided into 6 groups and then were put into 37 degree C water incubator for 0, 5, 10, 30, 60 and 120 minutes respectively. After washing, the target cells were fixed with Karnovsky's fixative and embedded in Epon 812. There were four intracellular routings for HAb 23, HAb18 and HAb8 to enter the QGY-7703 and SMMC-7721 hepatoma cells: (1) Coated pits coated: The colloid gold-antibodies which clustered in the specialised regions of the surface membrane were invaginated rapidly into the cells to form coated vesicles. (2) Enclosed invagination: One or two colloid gold-antibodies which were attached on the surface membrane were invaginated into the cell by endocytosis. (3) Microvilli involved routing: The microvilli which were adsorbed by the colloid gold-antibodies were broken and then were phagocytized by the cells. (4) Routing via glycocalyx-like material: The glycocalyx-like material which was clustered by the colloid gold-antibodies was invaginated during endocytosis.     

Down-regulation of lung resistance related protein by RNA interference targeting survivin induces the reversal of chemoresistances in hepatocellular carcinoma

Background Both survivin and lung resistance related protein （LRP） are hepatocellular carcinoma （HCC）. But the relationship between survivin and LRP is investigate the effects of down-regulation of survivin on LRP expressions and the both in vitro and in vivo. related to the chemoresistances in ndefinite. The aim of this study was to reversal of chemoresistances in HCC Methods The expressions of survivin were detected by RT-PCR and Western blotting in HCC cell line SMMC-7721 and SMMC-7721/ADM. The sensitivities of these two cell lines to ADM were evaluated by MTT assays. SiRNA which targeted survivin was transfected into SMMC-7721/ADM cells, then the sensitivity of SMMC-7721/ADM cells to ADM and the expressions of survivin and LRP were detected respectively. SMMC-7721/ADM cells were transplanted subcutaneously into nude mice to establish xenograft tumors. Antitumor activities of RNA interference （RNAi） targeting survivin, various doses of ADM and combination therapies were observed respectively. Possible toxicities were evaluated. LRP expression changes were tested. Student＇s ttest was used for evaluating statistical significance. Results The expressions of survivin in SMMC-7721/ADM cell line showed significant elevation compared to those in SMMC-7721 cell line （P 〈0.05）. Positive siRNA down-regulated the expressions of survivin significantly （P 〈0.05）. SiRNA targeting survivin could sensitize SMMC-7721/ADM cells to ADM and down-regulate the expressions of LRP significantly （P 〈0.05）. Growths of the tumors were significantly inhibited in positive siRNA group as compared with those in the control group from the 8th day （P 〈0.05）. Combination therapies caused significant tumor inhibitions compared with tumors of nude mice in the other three groups respectively （P 〈0.05）. No toxicities were found in nude mice treated by siRNA and combination therapies. The expressions of LRP were markedly reduced in tumors treated with siRNA targeting survivin （P 〈0.05）. Conclusions Down regulation of survivin gene by RNAi can increase chemosensitivity of HCC both in vitro and in vivo. The reversal of drug resistance may be reduced through the inhibitions of LRP.     

Internalization of immunotargeting drugs against hepatoma

Theeffectiveinternalizationoftheimmunotargetingdrugsisthepremiseforthemtodevelopaneffectiveimmunotargetingtreatment.Inrecentyears,scholarsbothhomeandabroadhaveaffirmedthecorrespondingprocessofinternalizationinthecelllinesofmelanoma,leukemia,gastriccarcinoma,etcLI'Zj,butuptonowtheproblemoftheinternalizationofimmunotargetingdrugsagainsthepatomahasnotyetbeenfullyunderstood.WeconjugatedADMparticleswiththemonoclonalantibodiesagainsthumanhepatocellularcarcinoma(HAb18andHAb25)byindirectconjugatin…     

Janus激酶抑制剂AG490对人肝癌SMMC-7721细胞侵袭转移的影响

目的 研究Janus激酶抑制剂AG490对人肝癌SMMC-7721细胞侵袭转移能力的影响,探讨JAK2/STAT3信号通路在肝癌侵袭调控中的作用。方法实验分为对照组、实验组（5、10 μmol/L AG490处理的SMMC-7721细胞）。采用Transwell小室检测肝癌细胞的侵袭能力,RT-PCR检测MMP-2 m...     

磁性白蛋白纳米球作为基因载体的研究

目的 制备载基因的磁性白蛋白纳米球,评估磁性白蛋白纳米球作为基因载体的可行性及体外的磁靶向性。方法 采用去溶剂化-交联法制备载基因磁性白蛋白纳米球,分别运用透射电镜（TEM）、动态光散射分析（DLS）对其形态、粒径进行表征。应用绿色荧光蛋白基因系统（pGFP）,观察载基因磁性白蛋白纳米球体外释放基因速率的情况及转染细胞的情况。运用普鲁士蓝染色法观察磁性纳米球的体外磁靶向性。结果 载基因磁性纳米球在TEM检测下为球形,大小均匀,并且在载基因磁性纳米球中明显地观察到在电子密度较低的白蛋白纳米球中包裹着电子密度较高的磁性纳米粒。DLS显示载基因免疫磁性纳米球的水合粒径约为209nm。体外动态释放基因速率,计算其累积释放率显示,该材料13h释放基因为95.25%,曲线平缓;转染实验结果显示载基因磁性纳米球能够有效转染SMMC-7721细胞,且转染效率高于载基因纳米球。普鲁士蓝染色实验结果显示磁靶向组细胞内的铁颗粒明显多于非靶向组。结论 成功制备了载基因磁性白蛋白纳米球,具有缓释基因的作用,并且能高效转染人肝癌细胞SMMC-7721。磁性白蛋白纳米球基因载体具有体外靶向性,有望作为一种潜在的靶向基因载体应用到生物医学领域。     

单纯疱疹病毒Ⅱ型胸苷激酶基因旁观者效应的实验研究

目的 :研究单纯疱疹病毒Ⅱ型胸苷激酶基因 (HSV -Ⅱtk)对人原发性肝细胞癌SMMC - 772 1的旁观者效应 .方法 :HSV -Ⅱtk基因通过阳离子脂质体Lipofectin介导 ,体外转染人原发性肝细胞癌SMMC -772 1,孵育 18h .取出细胞及上清液 ,对倍稀释 ( 2× ,4× ,8× ,16× ,3 2× )后分别加入另一 2 4孔板 .结果 :正常SMMC - 772 1细胞的浓度升高 ,亲本细胞的存活率随之降低 ,两者呈明显的负向相关关系 ;细胞组的OD值明显低于上清液组及空白对照组 (P <0 0 1) ;细胞组可见凋亡峰出现 ,而上清液组则无凋亡峰出现 .结论 :HSV -Ⅱtk基因对人原发性肝细胞癌SMMC - 772 1细胞具有旁观者效应 ,其作用机制可能与细胞紧密连接及细胞凋亡有关