热疗联合柔红霉素对CD34+CD38-白血病细胞增殖和凋亡的影响

目的 观察热疗联合柔红霉素(DNR)对CD34+ CD38-急性髓白血病KG1a细胞增殖的抑制作用和对其凋亡的影响.方法 免疫磁珠分离KG1a细胞中的CD34+ CD38- 细胞,以DNR作用48h后抑制50%CD34+ CD38- 细胞生长的药物浓度(IC50)作为后续实验的工作浓度.将CD34+CD38- 细胞分为对照组、热疗组(40℃、42℃)、化疗组和热化疗组(40℃、42℃).热疗、热化疗组在恒温水浴箱(40℃或42℃)hu加热60min,化疗组、热化疗组以IC50浓度的DNR处理48h.MTT法检测DNR对CD34+CD38- KG1a细胞增殖的抑制作用;流式细胞仪检测细胞凋亡率及42℃条件下细胞内DNR平均荧光强度的变化;甲基纤维素培养体系分析42℃条件下细胞的克隆形成能力.结果 KG1a细胞中分离出的CD34+CD38- 细胞纯度达95%以上,DNR的IC50为0.40μg/ml.热疗组、化疗组、热化疗组CD34+CD38-细胞增殖均明显受抑,且抑制作用随温度升高而增强(P<0.05).热疗组、化疗组及热化疗组细胞凋亡率均较对照组显著增加,且随温度升高凋亡率增加(P<0.05),热化疗组较化疗组及相同温度热疗组的细胞凋亡率显著增加(P<0.05).42℃热化疗组细胞内的DNR荧光强度(6.74±0.58)明显高于对照组(0.26±0.05)及化疗组(2.08±0.14,P<0.05).42℃热疗组、化疗组、42℃热化疗组细胞的集落形成率明显低于对照组,且42℃热化疗组明显低于42℃热疗组和化疗组(P<0.05).结论 热疗能增强DNR抑制CD34+ CD38- KG1a细胞增殖的作用,使细胞凋亡增多.     

槲皮素对人肝癌细胞SMMC7721增殖的影响

目的：研究槲皮素人肝癌细胞SMMC7721生长抑制的影响。方法：采用^3H-TdR掺入法,MTT检测法和双层琼脂集落形成实验,观察了肝癌细胞的生长变化。结果：槲皮素对人肝癌细胞SMMC77231有显的抑制生长及增殖,并能使肝癌细胞出现脱落死亡,DNA生成物减少,肝癌细胞SMMC7721集落形成率降低。结论：本实验研究提示,槲皮素对肝癌细胞SMMC7721具有较强的抑制生长及增殖性,并可能用于对肿瘤的协同的治疗研究。     

肝细胞肝癌组织及细胞株中PTPeta的表达及意义研究

目的 检测人肝细胞肝癌组织及细胞株SMMC7721中蛋白酪氨酸磷酸酶eta(PTPeta)的表达,观察肿瘤细胞生长密度对其表达的影响.方法 采用免疫组化方法检测肝细胞肝癌组织及SMMC7721细胞中PTPeta蛋白的表达;采用RT-PCR法检测不同生长密度(1×103、5×103、l×104、5×104/cm2)SMMC7721细胞中PTPeta的表达变化情况.结果 免疫组化检测结果表明肝细胞肝癌组织中PTPeta表达呈阳性,SMMC7721细胞中存在PTPeta表达,且定位于细胞膜;RT-PCR检测结果显示,当细胞密度为l×103、5×103、l×104、5×104/cm2时,PTPeta mRNA的表达量分别为0.425±0.031、0.659±0.041、0.771±0.043、0.885±0.045,PTPeta表达随着肿瘤细胞生长密度的增高而增加,差异具有统计学意义(P＜0.05).结论 PTPeta在肝细胞肝癌组织及细胞中表达,且随肿瘤细胞生长密度增高而表达增加.PTPeta 可能通过增加肝癌细胞黏附、维持恶性肿瘤细胞内环境稳定等,在恶性肿瘤细胞增殖及播散转移的过程中起重要作用.     

热疗联合榄香烯对SMMC 7721肝癌细胞HSP70的影响

目的探讨榄香烯联合热疗对肝癌细胞系SMMC 7721热休克蛋白70(HSP70)表达的变化以及与细胞凋亡的关系。方法采用肝癌细胞系SMMC 7721体外培养,MTT法测定单纯热疗、单纯榄香烯及榄香烯联合热疗对细胞增殖的影响。另外用流式细胞术的方法测定细胞凋亡与HSP70的表达。结果经热处理、榄香烯处理及联合处理,对肝癌细胞的生长均有明显的抑制作用,并诱导产生细胞凋亡。单纯热疗与单纯榄香烯处理细胞后HSP70表达增加,而榄香烯联合热疗后HSP70表达与前两组有所下降,细胞凋亡率却有所增加,与前两组比较差异有统计学意义(P<0.01)。结论榄香烯联合热疗下调了HSP70的表达,增加了细胞的凋亡率。     

纯化后海藻硫酸多糖-蛋白复合物诱导人肝癌细胞SMMC- 7721 凋亡作用的研究

目的 研究海藻硫酸多糖蛋白复合物(Sulfated Polysaccharide-Protein Complex,SPPC)诱导肿瘤细胞凋亡的效率和机制.方法 选用人肝癌细胞株SMMC-7721为实验对象,用MTT法,流式细胞术,DNA凝胶电泳法,细胞形态学检测方法对纯化后不同剂量SPPC处理的SMMC-7721细胞凋亡情况进行观察分析;并用Western blot 法检测凋亡相关蛋白 procaspase-3的表达量变化.结果 纯化后SPPC中、高剂量组(2.0、10.0mg · ml-1)可明显诱导SMMC-7721细胞凋亡, SMMC-7721细胞内凋亡相关蛋白procaspase-3有不同程度的表达降低,DNA凝胶电泳法检测到DNA ladder现象,流式细胞检测显示纯化后SPPC在10.0mg · ml-1浓度下出现了典型的二倍体峰.结论 一定浓度的海藻硫酸多糖蛋白复合物有诱导SMMC-7721细胞凋亡的作用,其生化机制可能是通过caspase-3 的活化来实现的.     

角燕提取物抑制肝癌生长的实验研究

目的：观察深海鱼类角燕提取物对肝癌的抑制作用。方法：采用盐溶液抽提，有机溶剂分级沉淀，分子筛层析等步骤，从海洋鱼类一角燕的药用部位获得角燕提取物。选用QGY7721人肝癌细胞体外实验模型，MTT法观察角燕提取物在体外对肝癌细胞生长的抑制作用；选用小鼠HAC肝癌模型，评价角燕提取物在体内对肝癌生长的抑制作用。结果：角燕提取物显著抑制QGY7721人肝癌细胞的生长，呈明显的剂量依赖关系，但对人肺成纤维细胞（HLF）的生长无明显影响。角燕提取物在剂量为0.25-1.00g/kg的范围内对小鼠HAC肝癌的生长均具有显著的抑制作用，并呈较好的剂量依赖关系；在剂量为1.00g/kg地，对小鼠HAC肝癌的抑瘤率为55．50％。结论：角燕提取物在体内和体外均能有效地抑制肝癌的生长。     

雌激素及其受体抑制剂在体外对胃癌细胞增殖的影响

笔者采用免疫荧光法检测证实人胃低分化腺癌SGC-7901细胞具有雌激素受体。在细胞培养过程中予以不同浓度的雌二醇处理,结果显示于10~(-6)—10~(-9)mol/L浓度时可刺激SGC-7901细胞的生长,以10~(-7)mol/L刺激作用最强,细胞群体倍增时间短于对照组(P<0.05),克隆形成率明显高于对照组(P<0.01),     

中药淡豆豉提取物的体外抗肿瘤作用研究

目的 研究中药淡豆豉提取物的抗肿瘤作用。方法 采用稻瘟霉分生孢子法初筛，四唑盐(MTT)比色法研究中药淡豆豉醇提物(SAE)对人肝癌细胞株SMMC-7721和QSG-7701生长的影响，并与其原料黑豆醇提物(HAE)作对比。结果 SAE可显抑制7721和7701生长，并且具有一定的时间、剂量依赖关系，并且作用强于HAE。结论 SAE体外具有抗肝癌细胞作用。     

红藤脂酸钠对人肝癌细胞系SMMC-7721体外抑瘤作用的实验研究

目的研究中药制剂红藤脂酸钠在体外对人肝癌细胞系SMMC-7721的生长抑制作用并初步探讨其作用机制。方法MTT法测定SMMC-7721细胞生长的抑制百分率,并运用AnnexinV法和TUNEL法检测凋亡发生率。结果MTT法测得对照组与红藤脂酸钠处理组吸光值(A值)分别为:对照组(0.385±0.03),红藤脂酸钠处理组中的250μg/ml组(0.31±0.019)、500μg/ml组(0.199±0.022)、1mg/ml组(0.15±0.033)和2mg/ml组(0.048±0.026);各组间比较有显著性差异(P<0.01)。AnnexinV法测定红藤脂酸钠(1mg/ml作用1h)处理后细胞凋亡90%,对照组细胞凋亡17.36%。TUNEL法测定红藤脂酸钠(1mg/ml作用1h)处理后细胞凋亡(60.3±6.0)%,与对照组细胞凋亡(6.6±3.5)%相比,差异有显著性(P<0.01)。结论红藤脂酸钠在体外对7721细胞生长有明显的抑制作用,其作用机制可能主要是诱导肿瘤细胞发生凋亡。     

热疗联合化疗对人胃癌活体细胞的抑制作用研究

目的探讨43℃热疗与常规化疗药物5-氟尿嘧啶(5-FU)等联合作用于人胃癌活体细胞时,何时进行热疗疗效最好,从而为临床治疗提供依据。方法利用MTT法测定43℃不同时间结合点热化疗组与单纯化疗组对人胃癌活体细胞的抑制率。结果43℃热疗与常规化疗药物5-FU等联合作用时,对人胃癌活体细胞的抑制率优于单纯化疗组(P<0.01)。结论43℃热疗与常规化疗药物5-FU等联合作用对人胃癌活体细胞的毒性作用更强。     

大黄素甲醚调节miR-370诱导肝癌细胞凋亡的实验研究

目的：观察大黄素甲醚调节miR-370诱导肝细胞癌（HCC）细胞的凋亡情况,并探讨其作用机制。 方法：将大黄素甲醚作用于SMMC7721和HepG2两组HCC细胞,使用TaqMan探针用实时定量PCR法检测miR-370的表达;用Western blot检测Sp1和DNMT1相应的蛋白表达水平。 结果：大黄素甲醚抑制肝细胞癌细胞增长和诱导凋亡。肝细胞癌细胞中通过上调miR-370造成大黄素甲醚诱导型细胞凋亡。大黄素甲醚处理的细胞中通过miR-370抑制剂抑制miR-370水平能明显降低凋亡细胞的百分比（P<0.01）。大黄素甲醚通过AMPK/Sp1/DNMT1信号调节miR-370的水平（P<0.01）。AMPK/Sp1/DNMT1信号参与大黄素甲醚诱导的HCC细胞凋亡。 结论：大黄素甲醚通过上调miR-370诱导HCC细胞凋亡,通过调节AMPK/Sp1/DNMT1信号通路对miR-370发挥调节作用。     

天冬多糖的提取及其对人肝癌SMMC-7721细胞生长影响的研究

目的 探讨天冬多糖提取方法的可靠性及天冬多糖对体外培养的人肝癌SMMC-7721细胞株生长的影响.方法 冷水浸提、乙醇沉淀法提取天冬粗多糖,再用链菌蛋白酶E及Sevag法脱蛋白,最后用纤维素及琼脂糖凝胶层析柱纯化提取纯品天冬脱蛋白多糖,并对其进行鉴定及分子量检测.用噻唑蓝法检测不同浓度纯品天冬脱蛋白多糖对于体外培养的人...     

乳腺癌缺失基因1对肝癌生物学行为的影响

目的探讨乳腺癌缺失基因1（deleted in breast cancer 1,DBC1）对肝癌细胞增殖和凋亡的影响。方法构建pcDNA3.1-DBC1的正义表达载体,转染肝癌细胞SMMC-7721,MTT法检测DBC1对肝癌细胞SMMC-7721生长增殖的影响,采用流式细胞术（FCM）检测DBC1对肝癌细胞SMMC-7721的细胞周期和细胞凋亡的影响。结果与转染空载体的SMMC-7721细胞相比,pcDNA3.1-DBC1正义表达载体转染上调DBC1的表达,可显著抑制肝癌细胞（SMMC-7721）增殖,升高G1期细胞比例（51.0%vs 64.9%,P=0.002）,降低S期细胞比例（29.7%vs 14.0%,P〈0.001）,诱导细胞凋亡（8.8%vs 24.6%,P〈0.001）。结论 DBC1通过抑制肿瘤细胞增殖,阻滞细胞周期进展,促进细胞凋亡,抑制肝癌的发生和发展。     

人前列腺癌细胞株PC-3中类肿瘤干细胞的培养分离和初步鉴定

目的 从人前列腺癌细胞系PC-3中分离前列腺癌类干细胞并进行初步鉴定,为前列腺癌干细胞的进一步研究奠定基础.方法分别用含血清及无血清培养基培养前列腺癌PC-3细胞,流式细胞仪检测两种不同培养条件下前列腺癌类干细胞的比例,细胞免疫荧光鉴定前列腺癌类干细胞,MTT法测定并比较前列腺癌类干细胞与前列腺癌PC-3细胞的增殖能力及倍增时间,观察前列腺癌类干细胞诱导分化过程及其分化后细胞的双重免疫荧光表达.结果 少量PC-3细胞能在添加了人表皮生长因子(EGF)、人碱性成纤维生长因子(bFGF)和人白血病抑制因子(LIF)的无血清培养基中存活并形成悬浮细胞球团,无血清培养基中前列腺癌类干细胞比例(1.43%)显著高于含血清培养基(0.41%,P<0.05).前列腺癌类干细胞的增殖能力(培养7d的细胞数为9.6×10~5个)强于前列腺癌细胞(培养7d的细胞数为3×10~5个,P<0.000 5),倍增时间(21.90h)短于前列腺癌细胞(28.38h,P<0.000 5),免疫荧光显示前列腺癌类干细胞表达CD133;前列腺癌类干细胞球可在含血清培养基中贴壁分化,且双重荧光检测显示其同时低表达CD44和CD133.结论 体外培养的前列腺癌细胞系PC-3中含有少量能表达前列腺癌干细胞表面标志物的前列腺癌类干细胞,并可通过无血清培养基富集这类类肿瘤干细胞.     

Selective inhibition of the epidermal growth factor receptor tyrosine kinase by BIBX1382BS and the improvement of growth delay, but not local control, after fractionated irradiation in human FaDu squamous cell carcinoma in the nude mouse

PURPOSE: To investigate the effect of BIBX1382BS, an inhibitor of the epidermal growth factor receptor tyrosine kinase, on proliferation and clonogenic cell survival of FaDu human squamous cell carcinoma in vitro, and on tumour growth and local tumour control after fractionated irradiation over 6 weeks in nude mice. FaDu human squamous cell carcinoma is epidermal growth factor receptor positive and significant repopulation during fractionated irradiation was demonstrated in previous experiments. MATERIALS AND METHODS: Receptor status, receptor phosphorylation, cell cycle distribution, cell proliferation and clonogenic cell survival after irradiation were assayed with and without BIBX1382BS (5 microM) in vitro. Tumour volume doubling time, BrdUrd and Ki67 labelling indices and apoptosis were investigated in unirradiated tumours growing in NMRI nude mice treated daily with BIBX1382BS (50 mg kg(-1) body weight orally) or carrier. Tumour growth delay and dose-response curves for local tumour control were determined after irradiation with 30 fractions within 6 weeks. RESULTS: BIBX1382BS blocked radiation-induced phosphorylation of the epidermal growth factor receptor and reduced the doubling time of FaDu cells growing in vitro by a factor of 4.9 (p=0.008). Radiosensitivity in vitro remained unchanged after incubation with BIBX1382BS for 3 days and decreased moderately after 6 days (p=0.001). BIBX1382BS significantly reduced the volume doubling time of established FaDu tumours in nude mice by factors of 2.6 when given over 15 days (p<0.001) and 3.7 when applied over 6 weeks (p<0.001). When given simultaneously to fractionated irradiation, growth delay was significantly prolonged by an average of 33 days (p=0.003). Local tumour control was not improved by BIBX1382BS. The radiation doses necessary to control 50% of the tumours locally were 63.6 Gy (95% confidence interval 55; 73) for irradiation alone and 67.8 Gy (60; 77) for the combined treatment (p=0.5). CONCLUSIONS: Despite clear antiproliferative activity in rapidly repopulating FaDu human squamous cell carcinoma and significantly increased tumour growth delay when combined with fractionated irradiation, local tumour control was not improved by BIBX1382BS. The results do not disprove that epidermal growth factor receptor inhibition might enhance the results of radiotherapy. However, the results imply that further preclinical investigations using relevant treatment schedules and appropriate endpoints are necessary to explore the mechanisms of action and efficacy of such combinations.     

索拉非尼联合奥沙利铂对肝癌HepG2细胞的抑制作用

目的 探讨索拉非尼联合奥沙利铂(L-OHP)对肝癌细胞HepG2的抑制作用及其可能的分子机制.方法 索拉非尼和L-OHP单独及联合给药后以CCK8法测定HepG2细胞的增殖,用流式细胞仪检测细胞周期及凋亡,用Western blot观察ERK及pERK蛋白的表达.结果 索拉非尼及L-OHP单药对HepG2均有抑制作用,...     

Selective inhibition of the epidermal growth factor receptor tyrosine kinase by BIBX1382BS and the improvement of growth delay,but not local control,after fractionated irradiation in human FaDu squamous cell carcinoma in the nude mouse

Purpose: To investigate the effect of BIBX1382BS, an inhibitor of the epidermal growth factor receptor tyrosine kinase, on proliferation and clonogenic cell survival of FaDu human squamous cell carcinoma in vitro, and on tumour growth and local tumour control after fractionated irradiation over 6 weeks in nude mice. FaDu human squamous cell carcinoma is epidermal growth factor receptor positive and significant repopulation during fractionated irradiation was demonstrated in previous experiments.

Materials and methods: Receptor status, receptor phosphorylation, cell cycle distribution, cell proliferation and clonogenic cell survival after irradiation were assayed with and without BIBX1382BS (5?µM) in vitro. Tumour volume doubling time, BrdUrd and Ki67 labelling indices and apoptosis were investigated in unirradiated tumours growing in NMRI nude mice treated daily with BIBX1382BS (50?mg?kg^?1 body weight orally) or carrier. Tumour growth delay and dose–response curves for local tumour control were determined after irradiation with 30 fractions within 6 weeks.

Results: BIBX1382BS blocked radiation‐induced phosphorylation of the epidermal growth factor receptor and reduced the doubling time of FaDu cells growing in vitro by a factor of 4.9 (p=0.008). Radiosensitivity in vitro remained unchanged after incubation with BIBX1382BS for 3 days and decreased moderately after 6 days (p=0.001). BIBX1382BS significantly reduced the volume doubling time of established FaDu tumours in nude mice by factors of 2.6 when given over 15 days (p<0.001) and 3.7 when applied over 6 weeks (p<0.001). When given simultaneously to fractionated irradiation, growth delay was significantly prolonged by an average of 33 days (p=0.003). Local tumour control was not improved by BIBX1382BS. The radiation doses necessary to control 50% of the tumours locally were 63.6?Gy (95% confidence interval 55; 73) for irradiation alone and 67.8?Gy (60; 77) for the combined treatment (p=0.5).

Conclusions: Despite clear antiproliferative activity in rapidly repopulating FaDu human squamous cell carcinoma and significantly increased tumour growth delay when combined with fractionated irradiation, local tumour control was not improved by BIBX1382BS. The results do not disprove that epidermal growth factor receptor inhibition might enhance the results of radiotherapy. However, the results imply that further preclinical investigations using relevant treatment schedules and appropriate endpoints are necessary to explore the mechanisms of action and efficacy of such combinations.     

青蒿素及其衍生物体外抗肿瘤活性研究

目的研究青蒿素及其衍生物体外抗肿瘤活性。方法采用四甲基偶氮唑盐法测定青蒿素、双氢青蒿素、蒿甲醚和青蒿琥酯体外抑制人肺癌细胞、人胃癌细胞、人结肠癌细胞、人红白血病细胞和人肝癌细胞的活性。结果青蒿素、双氢青蒿素、蒿甲醚和青蒿琥酯对5株肿瘤细胞有选择性的抑制作用。结论青蒿素及其衍生物在体外对不同的肿瘤细胞有抑制活性。