特素对胃癌细胞体外增殖抑制的实验研究

目的 探讨特素 (紫杉醇注射液 )在体外对人胃腺癌细胞的增殖抑制效应。方法 将不同浓度的特素与三株不同分化程度的人胃腺癌细胞株共同温育 ,采用体外药物敏感试验及流式细胞仪技术 ,检测特素对胃癌细胞的杀伤率 ,并对细胞周期的改变进行初步分析。结果 特素对三株胃癌细胞均存在显著的增殖抑制作用 ,对不同胃癌细胞的增殖周期影响不一。结论 特素体外对人胃癌细胞有良好的杀伤作用。

采用外周血单个核细胞体外增殖抑制试验预测突发性聋听力预后的价值

目的分析外周血单个核细胞体外增殖抑制试验预测突发性聋患者听力预后价值。方法将71例突发性耳聋患者根据入组前是否已接受过治疗分为初治组(41例)与难治组(30例),均行GC治疗,难治组在GC治疗基础上给予辅助治疗用药。两组入组前均实施PBMC体外增殖抑制试验,比较两组患者PBMC增殖抑制率的差别及听力预后。结果初治组PBMC增殖抑制率高于难治组,且初治组治疗后受损频率PTA低于难治组(P<0.05);SSNHL患者PTA改善与PBMC增殖抑制之间呈正相关性,患者的PTA改善值与发病至就诊时间之间呈负相关性。结论 PBMC的体外增殖抑制试验可以预测SSNHL患者对GC的敏感性,为早期制定科学、个体化的治疗方案打下基础。

平肺口服液成药及其含药血清体外抗肿瘤作用的比较

目的:通过观察中药复方平肺口服液成药及其含药血清在体外的抗肿瘤作用,探讨中药直接添加法和血清药理学方法的一致性。方法:中药复方直接添加法及血清药理学方法观察平肺口服液对人肺腺癌A549细胞增殖作用的影响,CCK-8法检测细胞增殖情况。结果:平肺口服液5 mg/mL和10 mg/mL在作用24、48和72 h均对A549细胞增殖存在影响,表现为生长抑制作用,与空白对照组比较,差异有统计学意义(P<0.05);在常规培养体系中添加20%含药大鼠血清培养至48 h时即对A549细胞增殖存在生长抑制作用,与空白对照组比较,差异有统计学意义(P<0.05),这种抑制作用随时间增长而增强。结论:中药复方平肺口服液成药及含药血清均具有抗肿瘤活性。

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles

AIM:To synthesize antisense oligonucleotides (ASODNs) of midkine (MK), package the ASODNs with nanoparticles, and to inhibit hepatocellular carcinoma (HCC) growth using these nanoparticles.METHODS: HepG2 cell proliferation was analyzed in vitro using the 3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt assay. The in vivo activity of nanoparticles delivering the MK-ASODNs was analyzed by histopathological and immunohistochemical staining and quantitative real time polymerase chain reaction (PCR). RESULTS: The in vitro proliferation of HepG2 cells was signif icantly inhibited by the nanoparticles packaged with MK-ASODNs (NANO-ASODNs). Furthermore, the NANO- ASODNs signif icantly inhibited the growth of HCC in the mouse model. CONCLUSION: NANO-ASODNs can significantly suppress the growth of HCC in vitro and in vivo.

Proliferation inhibition of human cervical cancer HeLa cells by Casticin in vitro

Objective： The aim of the study was to investigate the effect of Casticin on proliferation inhibition of human cervical cancer HeLa cells in vitro and to unravel the associated mechanisms. Methods： Human cervical HeLa cells were cultured in vitro. The inhibitory effect of Casticin on the viability of human cervical cancer HeLa ceils was evaluated by the MTT assay. The colony formation ability was detected by plate colony formation assay. Distribution of cell cycle was analyzed by flow cytometry. The protein expression levels were analyzed by Western blot. Results： Casticin significantly inhibited the growth of human cervical cancer HeLa cells in a dose- and time-dependent manner, and the IC50 was 2.82 μg/mL. The colony-forming rate was reduced drastically compared with control group （P 〈 0.05）. The cells were markedly arrested at G2/M phase after the treatment of Casticin for 48 h. Western blot showed that the expression of p21 protein was up-regulated and protein level of Cyctin B1 was depressed by Casticin in a concentration dependent manner. Conclusion： Casticin could inhibit the cell growth and lead to cell arrest in human cervical cancer HeLa cells, and the down-regulation of Cyclin B1 protein expression and activation of p21 protein might contribute to Casticin induced cell arrest in human cervical cancer HeLa cells.

The inhibiting effects of Laggera alata flavone on human ovarian cancer HO-8910 cells proliferation and its mechanism in vitro

The purpose of this study was to investigate the effect of Laggera alata flavonen （LAF） on the inhibit- ing effect of human ovarian cancer HO-8910 cells proliferation and its possible mechanism in vitro. Methods： Human ovarian cancer HO-8910 cells were cultured in vitro. Inhibitory effect of LAF on the viability of HO-8910 cells was evaluated by the MTT assay. Apoptotic effect of different concentrations of LAF on HO-8910 cells was assessed by AO/EB staining and FCM with propidium iodide （PI） staining. Expression of proteins related to apoptosis was analyzed by Western blot. Results： LAF significantly inhibited the viability of HO-8910 cells proliferation in a dose-dependent and time-dependent manner, there were statistical significance compared with NS group （P 〈 0.05）, and the ICso was 4.28 pg/mL for 48 h. The cells treated with LAF showed typical morphological change and apoptotic rate increased by FCM in a dose-dependent, and there was notable dif- ference compared with NS group （P 〈 0.05）. Western blot showed that expression of Fas, caspase-8, tBid and Cyto-c proteins were up-regulated after treatment with LAF for 48 h in a concentration dependent. Conclusion： LAF could inhibit HO-8910 cells proliferation and induce apoptosis, which may be through the pathway of death receptor in vitro.

三叶青水提物及其含药血清对人胃腺癌细胞株的抑制作用

目的:考察三叶青水提物及其含药血清对人胃腺癌细胞AGS体外增殖的影响。方法:清洁级小鼠随机分为5组:三叶青水提物高、中、低(20,10,5 g.kg-1)剂量组、环磷酰胺阳性对照(CTX,5 g.kg-1)组及阴性对照组(蒸馏水)。各组均ig给药,每次20 mL.kg-1,每天2次,连续给药3 d,最后1次给药后1 h断头取血,4℃静置4 h,3 000 r.min-1离心20 min,分离血清,56℃灭活30 min后,用0.22μm的滤膜过滤除菌,-20℃保存备用。对数生长期的AGS细胞经0.25%胰蛋白酶消化后,配成2.5×104/L的细胞悬液接种于96孔培养板,培养24 h后,加入不同浓度的三叶青水提物溶液及其含药血清,作用48 h后,采用MTT比色法在570 nm波长下测定A,观察不同浓度三叶青水提物及其含药血清对AGS细胞的抑制作用。结果:1～40 g.L-1三叶青水提物对AGS细胞有显著的抑制作用(P<0.01),IC50为13.15 g.L-1;三叶青水提物含药血清高、中、低(20,10,5 g.kg-1)剂量组也可显著地抑制AGS肿瘤细胞的增殖(P<0.01),IC50为70.06 g.L-1。二者均呈现出剂量依赖性。结论:三叶青水提物及其含药血清对AGS细胞均有较强抑制作用。

腺病毒介导反义c-myc增强骨肉瘤细胞对顺铂化疗敏感性的实验研究

目的构建表达反义cmyc的重组腺病毒,探讨重组腺病毒介导反义cmyc转染的人骨肉瘤MG63细胞对顺铂化疗敏感性的影响。方法应用基因重组技术,将约720bp的人cmyccDNA反向克隆到腺病毒载体,经重组、扩增、病毒包装后构建表达反义cmyc的重组腺病毒(AdAscmyc),并在体外转染骨肉瘤MG63细胞,采用瑞士染色、吖啶橙染色、蛋白免疫印迹(WesternBlot)、细胞体外增殖抑制试验(MTT)、流式细胞仪(FCM)等观察细胞形态、检测cmyc蛋白表达、瘤细胞体外增殖抑制、凋亡及细胞周期,分析AdAscmyc体外转染的骨肉瘤MG63细胞对顺铂化疗敏感性。结果成功构建AdAscmyc,滴度可达2×109pfu/ml,体外转染MG63细胞48h后,可降低cmyc蛋白表达,并与浓度为2.0、5.0μg/ml的顺铂作用2h后,可抑制MG63细胞的体外增殖,抑制率分为33.4%、54.2%,与对照腺病毒(AdLacZ)转染组相比差异有统计学意义(P<0.05),FCM检测证实AdAscmyc的转染可诱导骨肉瘤细胞凋亡,且顺铂治疗后凋亡比例增加,细胞周期分析显示AdAscmyc转染的骨肉瘤细胞出现G2/M期阻滞。结论腺病毒介导反义cmyc能诱导骨肉瘤MG63细胞凋亡并增加MG63细胞对顺铂化疗敏感性。

难治性突发性聋患者预后相关因素分析

目的:探讨糖皮质激素(GC)敏感性与难治性突发性聋(突聋)预后的关系,并分析影响难治性突聋预后的相关因素。方法:前瞻性对照观察91例难治性突聋患者,治疗前行外周单个核细胞体外增殖地塞米松(DEX)抑制实验,局部进行以甲泼尼龙为主的综合治疗,分析难治性突聋发病后3个月的疗效及影响预后的相关因素。结果:难治性突聋患者治疗总有效率为40.66%,性别、耳侧别、年龄、是否伴有眩晕或耳鸣、甲泼尼龙的局部使用方式与疗效无关,仅DEX抑制率、发病至就诊时间与疗效有关(R^2=0.712,0.118,均P<0.01),并且DEX抑制率与疗效相关性更强,GC有效组的DEX抑制率显著高于GC无效组。结论:DEX抑制率可用来反映难治性突聋患者的GC敏感性及预后,GC敏感性和发病至就诊时间是影响难治性突聋患者的2个重要因素,通过CCK-8法了解难治性突聋患者GC敏感性并尽早开始局部GC治疗,可改善预后。