缺氧诱导因子-1α(HIF-1α)反义寡核苷酸对宫颈癌细胞增殖的影响

目的研究缺氧诱导因子-1α反义寡核苷酸(asHIF-1α)对人宫颈癌裸鼠皮下移植瘤的抑制作用及其相关机理。方法使用宫颈癌细胞株He1a建立裸鼠皮下移植瘤模型,将荷瘤裸鼠随机分为3组,分别注射生理盐水,HIF-1α正义寡核苷酸和HIF-1α反义寡核苷酸,观察三组动物的肿瘤生长曲线,四周后处死裸鼠,计算肿瘤的体积及其抑制率,绘制皮下肿瘤生长曲线。应用免疫组化技术检测种植瘤内VEGF和HIF-1α的变化。结果 asHIF-1α有明显的抑瘤作用,肿瘤生长曲线减缓,体积抑制率为33%、瘤重抑制率为54%(P<0.05);反义治疗组种植瘤内HIF-1α和VEGF蛋白含量显著降低(P<0.05)。结论 asHIF-1α对实验动物荷瘤生长具有明显的抑制作用,其机制是通过抑制HIF-1α表达,进而抑制VEGF的合成,从而使肿瘤组织的血管形成被抑制。

Intervention of Mirtazapine on gemcitabine-induced mild cachexia in nude mice with pancreatic carcinoma xenografts

AIM: To investigate the effect of Mirtazapine on tumor growth, food intake, body weight, and nutritional status in gemcitabine-induced mild cachexia. METHODS: Fourteen mice with subcutaneous xenografts of a pancreatic cancer cell line (SW1990) were randomly divided into Mirtazapine and control groups. Either Mirtazapine (10 mg/kg) or saline solution was orally fed to the mice every day after tumor implantation. A model of mild cachexia was then established in both groups by intraperitoneal injection of Gemcitabine (50 mg/kg) 10 d, 13 d, and 16 d after tumor implanta- tion. Tumor size, food intake, body weight, and nutritional status were measured during the experiment. All mice were sacrificed at day 28. RESULTS: (1) After 7 d of gemcitabine administration, body-weight losses of 5%-7% which suggested mild cachexia were measured; (2) No significant difference in tumor size was detected between the Mirtazapine and control groups (P > 0.05); and (3) During the entire experimental period, food intake and body weight were slightly greater for the Mirtazapine group compared with controls (although these differences were not statistically significant). After 21 d, mice in the Mirtazapine group consumed significantly more food than control mice (3.95 ± 0.14 g vs 3.54 ± 0.10 g, P = 0.004). After 25 d, mice in the Mirtazapine group were also significantly heavier than control mice (17.24 ± 0.53 g vs 18.05 ± 0.68 g, P = 0.014). CONCLUSION: Mild cachexia model was successfully established by gemcitabine in pancreatic tumor-bearing mice. Mirtazapine can improve gemcitabine-induced mild cachexia in pancreatic tumor-bearing mice. It was believed to provide a potential therapeutic perspective for further studies on cachexia.

Study on Coagulation Activity of Callicarpa nudiflora

[Objective] This study aimed to study the coagulation effect of Callicarpa nudiflora. [Method] The effects of CaUtcarpa nudiflora on blood coagulation system of mice were investigated by measuring prothmmbin time （PT）, thrombin time（TT）, activated partial thromboplastin time （APTT） and plasma fibrinogen （FIB）, [Result] Compared with the control group, the crude extracts and ：eluted parts of Ca#/carpa nudiflora all significantly shortened PT, APTT and TT, and increased the content of FIB in mice, [Coadadon] The crude extracts and eluted parts of Callicarpa nudiflora have a coagulation-promoting effect through affecting several links of the coagulation process in mice,

Inhibitory effect of parvovirus H-1 on the formation of colonies of human hepatoma cell line in vitro and its tumors in nude mice

The inhibitory effect of parvovirus H-1 on the colonyforming ability in vitro of QGY-7703, a cultured human hepatoma cell line, and on the formation and growth of its tumors in nude mice was studied. With higher multiplicity of infection (MOI) of H-1 given, survival of the QGY-7703 cells was found to be decreased. H-1 DNA amplification level at 30 h postinfection(p.i.) was detected to be 7.4 times higher than that at 2 h by dispersed cells assay, while the cells were delayed to enter into S phase.Plaques were formed in the indicator cells (new-born human kidney cell line, NBK) by progeny H-1 virus particles released from the infected QGY-7703 cells by infectious cell center assay. The formation of tumors in nude mice by QGY-7703 cells which were injected s c at 2 h postinfection was observed to be prevented in 2 groups with given MOI 25 and 50. The tumor growth of MOI 10 group occurred at a lower exponential rate than that of control,after a 20 d latent period. It was evident that parvovirus H-1 exhibited a direct inhibitory effect on the formation and growth of human hepatoma cells in vivo as well as in vitro.

Bcl-2 Small Interfering RNA Inhibits the Growth of Human Lymphoma Transplanted Subcutaneously in Nude Mice

OBJECTIVE To investigate whether small hairpin RNA (shRNA)targeting Bcl-2 mRNA could inhibit the growth of lymphomatransplanted subcutaneously in nude mice.METHODS Recombinant Bcl-2 shRNA expression vector withgreen fluorescence protein (GFP) gene was constructed andpreserved in our lab. We evaluated the antitumor effect of the Bcl-2shRNA in vivo which was the model of nude mice bearing Rajicells xenografts. Human Raji cells were injected subcutaneouslyinto nude mice to establish lymphoma models. When thediameters of tumor were above 0.5 cm after Raji cells injection,the mice bearing tumor were randomly divided into four groups:saline control group, negative shRNA group, plasmid vectorgroup, Bcl-2 shRNA group. The polyethylenimine (PEI) was usedto transfect shRNA into tumor. The mixed PEI and shRNA wasinjected into tumors. The growth and size of tumor were observed.Tissue was stained by H&E for its pathological morphology. Theexpression of Bcl-2 mRNA in the tumor mass was detected byreverse transcription polymerase chain reaction (RT-PCR).RESULTS A significant difference in median tumor weightwas observed in mice treated with Bcl-2 shRNA, compared withthose in the groups of negative shRNA or plasmid vector or salinesolution (P< 0.05). Pathological evaluation was completed in allexcised tumors from nude mice bearing Raji cells xenografts.The tumor tissue of the mice treated with Bcl-2 shRNA showedapoptosis, serious necrosis of the cells and inflammatory cellsinfiltration. There was no change in the morphology of cellsamong negative shRNA, plasmid vector and saline solution group.In the group of the Bcl-2 shRNA, the expression levels of Bcl-2mRNA of the tumor tissue were effectively inhibited (P < 0.05).CONCLUSION The shRNA targeting at the Bcl-2 mRNAcould inhibit the growth of human lymphoma transplantedsubcutaneously in nude mice.

Jianpijiedu Fang improves survival of hepatocarcinoma mice by affecting phosphatase and tensin homolog, phosphoinositide 3-kinase, and focal adhesion kinase

OBJECTIVE: To investigate the effect of Jianpijiedu Fang (JPJDF) on phosphatase and tensin homolog (PTEN), phosphoinositide 3-kinase (PI3K), and focal adhesion kinase (FAK), and on the survival of hepatocellular carcinoma (HCC) nude mice. METHODS: Forty male nude mice were randomly divided into 4 groups. Human HCC tissue was implanted in the livers of three groups. After 24 h, the three groups were treated respectively with JPJDF (37.5 g/kg), saline (20 mL/kg) and Tegafur (FT-207, 160 mg/kg) once a day for 10 weeks. The control group without implanting the tissue was concurrently treated with saline (20 mL/kg). The survival data and body weight of all mice were recorded, and expression levels of PTEN, PI3K and FAK in normal tissue and cancer tissue of the livers were eval-uated with immunohistochemical method. RESULTS: The cumulative survival rate of the mice in the JPJDF group was higher than those of the other groups. The rate of weight loss was the lowest in JPJDF group. The survivability and weight loss rate in FT-207 group were the poorest in all groups. The expression intensity of PTEN was higher in normal tissues than in cancer tissues, and lower in the normal tissues of HCC models than in that of mice without HCC. The PTEN expression intensity in normal tissue and cancer tissue from mice treated with FT-207 were lower than that from the mice treated with JPJDF or saline.The expression intensity of PI3K was higher in cancer tissue than in normal tissue. The PI3K expression intensity was the lowest in normal tissue and cancer tissue from mice treated with JPJDF, and the intensity from mice treated with FT-207 was the highest. In mice treated with JPJDF, the expression intensity of FAK was higher in the normal tissue and lower in the cancer tissue than those of the other treatment groups. CONCLUSION: The mechanism accounting for the prolonged survival of HCC-bearing mice treated with JPJDF might be related to the reduction in weight loss and the benign regulation of PTEN, PI3K, and FAK.