受体外周血IgG、MΦ、NK、CD4、CD8及MLR的变化与异种移植物存活的关系

目的探讨受体外周血IgG、MΦ、NK、CD4、CD8及MLR的变化与异种移植物存活的关系。方法选用小鼠移植于大鼠耳后心肌组织模型,按实验所设Ⅰ、Ⅱ、Ⅲ、Ⅳ组在术前12、8、4、0d及10、6、2、0d分别将小鼠脾细胞1×10     

去甲斑蝥素对人胆囊癌侵袭转移的抑制作用及机制

目的：探讨去甲斑蝥素(norcantharidin,NCTD)对人原发性胆囊癌侵袭和转移的影响及其机制。方法：应用细胞培养技术培养人原发性胆囊癌GBC-SD细胞,通过建立裸鼠人胆囊癌移植瘤模型进行随机药物体内实验,实验分为空白(生理盐水)对照组、5-FU组、NCTD组、5-FU NCTD组4组,观察荷瘤裸鼠移植瘤的瘤周组织浸润和肺转移；同时应用免疫组化SABC法检测NCTD对移植瘤细胞中nm23、MMP-2、TIMP-2蛋白表达的影响。结果：在动物实验中,NCTD可明显减少荷瘤裸鼠移植瘤的瘤周组织浸润和肺转移瘤结节(P＜0.05)；免疫组化检测显示NCTD组MMP-2表达下降,nm23、TIMP-2表达上升(P＜0.05)；若与5-FU联合应用上述作用更明显。结论：在裸鼠人胆囊癌模型中,NCTD可明显抑制人胆囊癌的侵袭和转移,若与5-FU联合应用则具协同作用,其机制可能与NCTD影响基质溶解和转移相关基因表达有关。     

苏拉明联合顺铂对肺腺癌小鼠移植瘤生长和转移的影响

背景与目的:新近研究发现新生血管生成在肿瘤生长、转移以及转移灶的生长过程中起重要作用。本研究观察苏拉明联合顺铂对肺腺癌细胞LA795的T739小鼠异体移植瘤生长和转移的抑制作用,并初步探讨其作用机制。方法:建立肺腺癌细胞LA795的T739小鼠异体移植瘤模型,将32只接种LA795细胞T739小鼠随机分成4组,每组8只。对照组:每只小鼠每天生理盐水0.2ml腹腔注射;顺铂组:顺铂2mg·(kg·d)-1于接种后第4、11、18天各一次;苏拉明组:苏拉明10mg·(kg·d)-1;顺铂 苏拉明组:顺铂2mg·(kg·d)-1于接种后第4、11、18天各腹腔注射一次 苏拉明10mg·(kg·d)-1。用药16日,用药中观察肿瘤生长情况,于接种后第24天处死各组小鼠,取出双肺并剥离皮下肿瘤,计算出肺转移发生率,计数各组小鼠肺表面转移结节数并算出肺表面结节转移抑制率。收集移植瘤标本行光镜观察,采用免疫组化和图像分析系统定量检测肿瘤组织微血管密度(microvesseldensity,MVD),血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)及核因子-κB(nuclearfactor-kappaB,NF-κB)的表达和原位凋亡TUNEL法检测肿瘤细胞凋亡指数。结果:顺铂组、苏拉明组、顺铂加苏拉明组肿瘤的生长明显受到抑制,瘤重明显低于对照组,其抑瘤率分别为23.0%、34.4%、56.3%,而联合用药组抗瘤作用进一步增强。光镜观察顺铂组、苏拉明组、顺铂加苏拉明组肿瘤细胞出现坏死,苏拉明组和顺铂加苏拉明组肿瘤间质中血管数减少。各用药组NF-#B的表达都比对照组减少,凋亡指数比对照组明显提高,差异有显著性(P<0.01);苏拉明组及联合组与对照组和顺铂组相比肺表面转移结节数明显下降,同时肺转移发生率、皮下肿瘤MVD、VEGF的表达也明显下降,相反顺铂组对此则无明显改变。结论:苏拉明可明显抑制肺腺癌细胞在小鼠体内的生长和转移,与顺铂联用有协同作用,其作用机制可能与抑制其微血管形成、促进细胞凋亡有关。     

In vivo antitumor efficacy of 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride), a water-soluble geldanamycin derivative

Purpose To describe the preclinical basis for further development of 17-dimethyl aminoethylamino-17-demethoxygeldanamycin hydrochloride (17-DMAG, NSC 707545).Methods In vitro proliferation assays, and in vivo model studies in metastatic pancreatic carcinoma and subcutaneous xenograft melanoma and small-cell lung carcinoma models.Results 17-DMAG emerged from screening studies as a potent geldanamycin analog, with the average concentration inhibiting the growth of the NCI anticancer cell line drug screen by 50% being 0.053 M. Head to head comparison with 17-allylamino-17-demethoxygeldanamycin (17-AAG, NSC 330507) revealed 17-DMAG to possess potent activity against certain cell types, e.g., MDA-MB-231 breast carcinoma and HL60-TB leukemia which were relatively insensitive to 17-AAG. Evidence of oral bioavailability of 17-DMAG in a saline-based formulation prompted more detailed examination of its antitumor efficacy in vivo. 17-DMAG inhibited the growth of the AsPC-1 pancreatic carcinoma xenografts growing as intrahepatic metastases at doses of 6.7–10 mg/kg twice daily for 5 days administered orally under conditions where 17-AAG was without activity. 17-DMAG in an aqueous vehicle at 7.5–15 mg/kg per day for 3 days on days 1–3, 8–10 and 13–17, or 1–5 and 8–12 showed evidence of antitumor activity by the parenteral and oral routes in the MEXF 276 and MEXF 989 melanomas and by the parenteral route in the LXFA 629 and LXFS 650 adenocarcinoma and small-cell carcinoma models. The latter activity was comparable to the historical activity of 17-AAG.Conclusions Taken together, the in vivo activity of 17-DMAG supports the further development of this water-soluble and potentially orally administrable geldanamycin congener.     

Predictive impact of activated leukocyte cell adhesion molecule (ALCAM/CD166) in breast cancer

Accumulating evidence indicates that breast cancer is caused by cancer stem cells and cure of breast cancer requires eradication of breast cancer stem cells. Previous studies with leukemia stem cells have shown that NF-κB pathway is important for leukemia stem cell survival. In this study, by using MCF7 sphere cells as model of breast cancer stem-like cells, we evaluated the effect of NF-κB pathway specific inhibitors on human breast cancer MCF7 sphere cells. Three inhibitors including parthenolide (PTL), pyrrolidinedithiocarbamate (PDTC) and its analog diethyldithiocarbamate (DETC) were found to preferentially inhibit MCF7 sphere cell proliferation. These compounds also showed preferential inhibition in term of proliferation and colony formation on MCF7 side population (SP) cells, a small fraction of MCF7 cells known to enrich in breast cancer stem-like cells. The preferential inhibition effect of these compounds was due to inhibition of the NF-κB activity in both MCF7 sphere and MCF7 cells, with higher inhibition effect on MCF7 sphere cells than on MCF7 cells. PDTC was further evaluated in vivo and showed significant tumor growth inhibition alone but had better tumor growth inhibition in combination with paclitaxel in the mouse xenograft model than either PDTC or paclitaxel alone. This study suggests that breast cancer stem-like cells could be selectively inhibited by targeting signaling pathways important for breast cancer stem-like cells.     

In vivo antitumor activity by 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone in a solid human carcinoma xenograft model

Previously we have shown that 2,4-dihydroxy-6-methoxy-3,5-dimethylchalcone (DMC), which is isolated from the buds of Cleistocalyx operculatus, significantly inhibits the growth of human liver cancer SMMC-7721 cells and is able to induce apoptosis of SMMC-7721 cells in vitro. Here we report the antitumor effects of DMC in vivo, using a solid human tumor xenograft mouse model using human liver cancer SMMC-7721 cells. The average tumor weights in the control group and in mice injected with 150 mg/kg DMC were 1.42±0.11 g and 0.59±0.12 g, respectively. Flow cytometric analysis of the tumor cell population demonstrated an aneuploid peak (representing 33.60±0.80% of the total in mice injected with 150 mg/kg DMC). To our knowledge, this is the first time that chalcone compounds have been applied to a human tumor xenograft model.     

ILK-ASODN对人卵巢上皮癌裸鼠皮下移植瘤形成的抑制作用

目的 探讨ILK-ASODN对人卵巢上皮癌裸鼠皮下移植瘤形成的抑制作用。方法 研究分为对照组和实验组。在转染后72 h采用RT-PCR和Western blot法检测细胞ILK mRNA及蛋白表达量;运用流式细胞术评价ILK-ASODN转染对肿瘤细胞周期的影响;同时将转染后的HO-8910细胞接种于裸鼠皮下,观察裸鼠体重变化、体内成瘤率、肿瘤出现的时间、肿瘤体积及重量的变化,并计算抑瘤率。结果 实验组与对照组比较,细胞的ILK mRNA及蛋白表达量显著下降、G₀/G₁期细胞明显增多,差异具有统计学意义(P＜0.01);实验组裸鼠肿瘤出现时间明显晚于对照组,肿瘤体积及重量增长速度显著减缓,差异有显著统计学意义(P＜0.01)。结论 ASODN可以阻断人卵巢癌细胞ILK基因表达,抑制细胞生长;而 ILK基因的表达沉默对裸鼠皮下移植瘤的形成具有明显抑制作用。     

Enhanced therapeutic effect of cisplatin on the prostate cancer in tumor-bearing mice by transfecting the attenuated Salmonella carrying a plasmid co-expressing p53 gene and mdm2 siRNA

Prostate cancer urgently needs an efficient therapy. Here we demonstrated that cisplatin combined with gene therapy by transfecting the attenuated Salmonella that carry a plasmid containing p53 gene and MDM2 siRNA provided a super-synergistic effect on the inhibition of prostate cancer growth in vivo. This synergistic therapy was associated with the induction of apoptotic cell death with a decreased Bcl2 to Bax expression ratio and increased expression of cleaved caspase 3 and caspase 9 in the prostate cancer xenograft. These results indicate that cisplatin-chemotherapy in combination with targeting the MDM2/p53 axis is an attractive strategy to treat prostate cancer.     

CD19基因过表达K562细胞株及NOD-SCID小鼠皮下移植瘤模型的建立

目的 建立人CD19基因过表达的K562细胞株(CD19-K562)NOD-SCID小鼠皮下移植瘤模型.方法 克隆人CD19基因构建MigR1-CD19反转录病毒载体,转入Plat-A包装细胞,病毒上清转染K562细胞,反转录聚合酶链反应(PCR)及流式细胞术检测转染细胞CD19基因和蛋白表达,体外反复传代获得CD19-K562稳定转染细胞株,锥虫蓝染色,细胞计数检测细胞增殖,Annexin V/碘化丙啶(PI)双染流式细胞术检测细胞凋亡.CD19-K562细胞株皮下接种NOD-SCID小鼠并经体内外传代后建立移植瘤亚系CD19-K562-a,建立NOD-SCID小鼠皮下移植瘤模型.应用反转录PCR、瑞特染色、免疫组织化学等方法验证CD19-K562-a细胞性质.结果 成功建立了人CD19-K562细胞稳定株,CD19阳性率(99.80±0.17)％.CD19-K562细胞增殖、凋亡未受转染及传代影响.CD19-K562细胞稳定株皮下接种NOD-SCID小鼠后经体外结合体内传代建立移植瘤亚系CD19-K562-a,其CD19阳性率为(99.78±0.04)％,CD19-K562-a和CD19-K562细胞均呈未分化形态.CD19-K562-a皮下接种NOD-SCID小鼠成功建立移植瘤模型,CD19-K562-a瘤体CD19基因表达强阳性.结论 通过构建MigR1-CD19重组载体获得CD19基因过表达的K562稳定转染细胞株(CD19-K562),同时采用体外结合体内传代建立稳定成瘤的NOD-SCID小鼠移植瘤亚系CD 19-K562-a并成功建立NOD-SCID小鼠皮下移植瘤模型.     

靶向IGF1R的siRNA抑制人肝癌细胞SMMC7721裸鼠移植瘤的实验研究

背景与目的:利用小干扰RNA(small interfering RNA,siRNA)抑制哺乳动物基因表达已成为研究基因功能的一种有效方法.本研究探讨人胰岛素样生长因子1类受体(insulin-like growth factor 1 receptor,IGF1R)的短发夹环RNA对人肝癌细胞株SMMC7721裸鼠移植瘤的抑制作用.方法:设计并合成特异性靶向IGFIR的siRNA片段,构建SMMC7721-IGFlR-siRNA表达质粒,将其转入SMMC7721细胞.通过G418筛选出稳定株,移植裸鼠成瘤.同时设立对照组.根据体积的均数值绘制肿瘤生长曲线,以HE染色法观察质粒处理后瘤组织的病理改变,Western blot检测肿瘤组织中IGF1R的表达变化,免疫组化SP法检测检测瘤组织内微血管密度,原位末端标记法(TUNEL法)检测肿瘤细胞的凋亡情况.结果:SMMC7721-IGF1R-siRNA组肿瘤体积与SMMC7721-IGF1R-mutation组、SMMC7721组相比差异有统计学意义(P＜0.05).病理学检查及TUNEL检测发现,SMMC7721-IGF1R-siRNA组肿瘤生长受到抑制,细胞凋亡指数[(50.2±6.4)%]明显高于SMMC7721.IGF1R-mutation 组[(5.4±1.0)%]和SMMC7721组[(6.0±2.1)%](P＜0.05).SMMC7721-IGF1R-siRNA组与SMMC7721-IGF1R-mutation 组、SMMC7721相比,瘤内IGFIR蛋白表达明显下调.SMMC7721-IGF1R-siRNA组微血管密度(11.3±4.4)与SMMC7721-IGF1R-mutation组(36.7±7.6)、SMMC7721组(28.4±6.5)相比明显下降(P＜0.05).结论:构建的SMMC7721-IGF1R-siRNA具有RNA干扰作用,能抑制SMMC7721细胞裸鼠移植瘤的生长.