The model of transplanted colonic SW480 cell line carcinoma in gymnomouse body was set up to observe the effect of octapeptide somatostatin (SMS 201-995,SMS) on the transplanted carcinoma and elucidate its mechanism. Results: the volume, weight, DNA and protein content in carcinoma cell, cell amount and proliferation index of S and G2M phase in SMS group and SMS+PG (pentagastrin) group were markedly lower than those in PG group and control group, those of PG group were markedly higher than those in control group.The cell amount of G0/G1 phase in SMS group and SMS+PG group was markedly higher than that in PG group and control group, and that of PG group was markedly lower than that in control group.All these suggested that somatostatin could not only inhibit the growth of transplanted human colonic SW480 cell line carcinoma directly but also inhibit the growthpromoting effect of gastrin on the transplanted carcinoma.The mechanism might be that somatostatin inhibit the synthesis of cAMP, DNA and protein in carcinoma cells, then inhibit the cell growing from G0/G1 phase to S and G2M phases.Our study might provide experimental basis for the homonotherapy with analogue of somatostatin in patients with large intestine carcinoma.
目的 通过复制人肝癌细胞株HepG2裸鼠皮下移植瘤模型，观察绿茶提取物表没食子儿茶素没食子酸酯（EGCG）干预对HepG2移植瘤新生血管生成的影响。 方法 瘤体接种复制HepG2移植瘤模型，荷瘤裸鼠20只随机分组，实验组给予EGCG溶液每日20 mg/（kg·只），腹腔注射3周，对照组给予等量灭菌注射用水3周，末次用药24 h，后处死裸鼠，剥离移植瘤。常规病理切片观察移植瘤组织结构；逆转录-聚合酶链式反应和免疫组织化学法检测移植瘤缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）mRNA及蛋白表达，并通过检测CD34表达计数瘤组织微血管密度（MVD）。 结果 组织病理学观察实验组移植瘤见大量坏死区，瘤体内血管数量明显少于对照组；实验组HIF-1α、VEGF mRNA及蛋白表达水平比对照组均明显下调（P＜0.05），实验组MVD比对照组明显下降（P＜0.05）。 结论 EGCG可抑制荷瘤裸鼠HepG2移植瘤新生血管生成。
Objective To establish a xenograft model of hydroxycamptothecine (HCPT)-resistant human gastric cancer cell line (SGC-7901/HCPT) in nude mice and study its biological characteristics. Methods The SGC-7901 and SGC-7901/ HCPT cells were cultured in vitro. The cell suspension was injected subcutaneously into the nude mice. When the subcutaneous carcinoma was 1.0 cm in diameter, it was cut off and divided into pieces of 0.1-0.2 cm in diameter． Then the small pieces of tumor were re-transplanted subcutaneously into the second generation nude mice until the fourth generation. The morphological feature, ultramicro-structure, and growth characteristics of the fourth generation transplanted tumor were examined. The drug resistance was measured by methyl thiazolyl tetrazolium (MTT) assay. Results The transplanted tumor in nude mice was round or oval, and many blood vessels were on its surface. Under the light microscope, the sizes of SGC-7901 transplanted tumor cells were similar, and sizes of cell nuclei were also similar; Meanwhile, the morphous of SGC-7901/HCPT transplanted tumor cells were irregular and in disorder, and the size of the cell nuclei was different from each other. Under the electron microscope, the mitochondria and endoplasmic reticulum of SGC-7901 transplanted tumor cells were nearly normal and no swelling in cell nuclei; Meanwhile the cell nuclei of SGC-7901/HCPT transplanted tumor cells were lightly swelled, a the mitochondria and endoplasmic reticulum were obviously swelled. By MTT assay, compared with SGC-7901 transplanted tumor cells, the resistance index of SGC-7901/HCPT transplanted tumor cells was 9.02±0.78 in HCPT, and resistance index to Adriamycin， Mitomycin C, 5-fluorouracil, and Etoposide was 1.24±0.09, 1.31±0.17, 0.96±0.12, and 1.07±0.16, respectively. Conclusions A transplanted tumor model of SGC-7901/HCPT in nude mice is established successfully， and showing stable drug resistance to HCPT and no cross-resistance to other chemotherapeutics, which can be used for further experiments.
Objective To explore the effect on apoptotic genes of pancreatic adenocarcinoma cell BxPC-3 from subcutaneous transplantation tumor in nude mice induced by 5-FU and sulfasalazine (SZ).Methods Changes of apoptosis-related genes 〔bcl-2, cyclinD1, Bax and NF-κB (p65)〕 in subcutaneous transplantation tumor treated by 5-FU, SZ alone or both at the levels of mRNA and protein were measured by RT-PCR and Western blot. Results NF-κB (p65) at mRNA relative content and protein expression in subcutaneously heterotopic transplantation tumor treated by 5-FU (7.5, 15 mg/kg), SZ (10, 20 mg/kg) alone or both showed significant difference, except for two subsets in SZ group, respectively, in comparison with each control group (P＜0.01). Meanwhile bcl-2 and cyclinD1 at the levels of mRNA and protein, and Bax protein level were significantly different from each control group (P＜0.01). The above-mentioned indexes were show obvious interaction of both by multiple factor analysis of variance. Conclusion Up-regulated level of Bax, down-regulated levels of bcl-2, cyclinD1 and NF-κB (p65) might be one of apoptotic mechanisms that SZ synergistically enhanced apoptotic effect on pancreatic adenocarcinoma cell BxPC-3 of subcutaneous transplantation tumor in nude mice induced by 5-FU.
ObjectiveTo explore the effect of Poria cocos on xenograft tumors of gastric cancer SGC-7901 cell line in mude mice. Method①After establishment of xenograft tumor of gastric cancer SGC-7901 cell line, 10 nude mice were equally divided into normal control group and Poria cocos group. The nude mice of each group were gavaged with normal saline (NS) and Poria cocos (0.5 mL) for 32 days, respectively. Tumor volume were measured to draw tumor growth curves and the tumor weight inhibitory rate was calculated with tumor weight (on the 32-day, nude mice were sacrificed to get the xenograft tumors). The expressions of B cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), Caspase-3, Caspase-9, and vascular endothelial growth factor (VEGF) were detected by immunohistochemical staining. ②Preparation of drug serum containing Poria cocos. Gastric cancer SGC-7901 cell line were be divided into 2 groups: normal control group and Poria cocos group. Cells of normal control group were treated with serum containing NS, and cells of Poria cocos group were treated with drug serum containing 10% Poria cocos. After 24 hours and 48 hours, Western-blot was used to detect the expressions of Bcl-2 and Bax. ResultsOn 32-day, the volume and weight of xenograft tumors in normal control group〔(2 652.17±225.01) mm3 and (2.48±0.21) g〕were both higher than those of Poria cocos group〔(1 247.56±277.23) mm3 and (1.28±0.28) g〕, P<0.050. The tumor inhibitory rate in Poria cocos group was 48.39%. The results of immunohistochemical staining showed that, compared with normal control group, Poria cocos could down-regulate the expressions of Bcl-2〔(4.20±1.10）score vs. (8.00±1.20) score〕and VEGF〔(3.80±0.45) score vs. (7.80±1.10) score〕, while up-regulate the expressions of Bax〔(7.40±1.34) score vs. (3.00±0.71) score〕, Caspase-3〔(6.60±1.34) score vs. (2.60±0.55) score〕, and Caspase-9〔(7.20±1.79) score vs. (4.00±1.22) score〕, P<0.050. Compared with normal control group (1.72±0.03), the expression value of Bcl-2 was all higher in 24 h-Poria cocos group (0.96±0.04) and 48 h-Poria cocos group (0.77±0.04), P<0.050, and the expression value was higher in 48 h-Poria cocos group than that of 24 h-Poria cocos group (P<0.050). Compared with normal control group (0.15±0.01), the expression value of Bax was higher in 48 h-Poria cocos group (0.55±0.01), P<0.050, but there was no significant difference between the normal control group and 24 h-Poria cocos group（0.19±0）, P>0.050. ConclusionsPoria cocos can restrain the growth of xenograft tumors for gastric cancer SGC-7901 cell line in mude mice, and the mechanism may be related to mitochondrial apoptosis pathway and the inhibition of expression of VEGF.
ObjectiveTo observe the effects of hydroxysafflor yellow A (HSYA) on microvessel density (MVD) of mice transplanted Lewis lung cancer and mRNA expression of vascular endothelial growth factor (VEGF) so as to explore the tumor-inhibiting mechanism of HSYA. MethodsSixty tumor-bearing C57/BL mice were randomly divided into five groups, with 12 mice in each group, namely a control group, a cyclophosphamide (CTX) group (25mg/kg), a large dose HSYA group (112mg/L), a medium dose HSYA group (56mg/L), and a small dose HSYA group (28mg/L). These different drugs were administered by intraperitoneal injection. The mice were sacrificed 22 days after the treatment. Tumor tissues were sampled and examined by immunohistochemical method and quantitative real-time PCR to detect the expression of MVD and VEGF mRNA. ResultsThe MVD of the medium and small dose HSYA groups and CTX group were 30.01±3.12, 22.56±2.11 and 16.21±2.40, respectively, which were significantly lower than 41.10±2.93 of the control group and 37.66±3.04 of the large dose HSYA group (χ2=2.82, P=0.010;χ2=3.16, P=0.007;χ2=4.58, P=0.000) and (χ2=1.98, χ2=0.038;χ2=2.45, P=0.016;χ2=3.82, P=0.001). The difference in VEGF amplified fluorescence expression threshold between the HSYA groups and the control group was not significant. However, after amplification, the expression of VEGF mRNA in the small dose HSYA group was only 0.43±0.16, which was obviously lower than 0.82±0.06 in the control group (F=0.77, P=0.038). ConclusionHSYA can significantly reduce MVD in mice transplanted Lewis lung cancer and down-regulate expression of VEGF mRNA to achieve tumor-inhibiting effect.
Objective To observe the growth of xenografted tumor in nude mice after DDX46 expression decreased, and to further study the role of DDX46 in the development and progression of esophageal squamous cell carcinoma. Methods DDX46-shRNA mediated RNAi was applied to silencing DDX46 in Eca-109 cells. Twenty-five female BALB/c nude mice were divided into 3 groups: an experiment group (DDX46-shRNA-LV, n=10), a control group (Control-LV, n=10) and a blank control group (Het-1A, n=5). The prepared Eca-109 cells of DDX46-shRNA-LV and Control-LV were subcutaneously injected into the right armpit of mice (4×106 cells per mouse), while Het-1A cells were subcutaneously injected into the bilateral armpits of mice (4×106 cells per side). Tumor growth was monitored twice a week on the 14th day after injection. Tumor volume was measured with calipers, in vivo imager to observe the fluorescence of each group. Further, western blotting analysis was used to detect the changes of apoptosis signaling molecules in xenografted tumor after DDX46 silence. Results The growth of xenografted tumor in nude mice was significantly slower in the DDX46-shRNA-LV group than that in the Control-LV group throughout the study period (P<0.001). Western blotting analysis showed that silencing DDX46 effectively suppressed the expression of DDX46, and upregulated the expression of cleaved Caspase-3 and cleaved PARP-1 in xenografted tumor (P<0.01). Conclusion DDX46 is involved in the development and progression of esophageal squamous cell carcinoma, and the silence of DDX46 expression can inhibit the growth of esophageal squamous cell carcinoma, which probably by positive regulation of apoptosis signaling pathway.
ObjectiveTo optimize the culture method of human primary pancreatic ductal adenocarcinoma (PDAC) cells and cancer associated fibroblasts (CAFs) and investigate the effect of CAFs on the growth of primary PDAC cells in vitro and tumor formation in patient-derived xenograft (PDX) model.MethodsThe PDAC specimens were collected and primarily cultured. In order to observe the effect of CAFs on the growth of primary PDAC cells in vitro, the CAFs were co-cultured with primary PDAC cells consistently and the alone cultured primary PDAC cells served as the control. Then, these cells were injected into the shoulder blades of NOG mice in order to develop the PDX model.ResultsWhen the primary PDAC cells separated from the CAFs, the proliferation capacity of the primary PDAC decreased rapidly in the passage culture in vitro, and the most cells were terminated within 5 generations. By contrast, when the CAFs co-cultured with the primary PDAC cells, the proliferation capacity of primary PDAC cells were preserved, which could be stably transferred to at least 10 generations. The tumors of NOG mice were detected during 2–3 weeks after injecting the mixed cells (primary PDAC plus CAFs), while had no tumor formation after injecting CAFs alone. The rate of tumor was 92.9% (13 cases) in the primary PDAC plus CAFs group, which was higher than that of the CAFs alone group (64.3%, 9 cases), but there was no statistical difference because of the small sample size. The volume of tumor in the primary PDAC plus CAFs group at 2, 4, 6, and 8 weeks after the tumor cells injection was significantly larger than that in the CAFs alone group at the corresponding time point, the differences were statistically significant (P<0.01).ConclusionsThe CAFs could promote the growth of primary PDAC cells in vitro. This new method of co-culture CAFs with primary PDAC could improve the success rate of primary PDAC cells culture and improve the success rate of PDX model in NOG mice.
ObjectiveTo investigate the inhibitory effect of short hairpin RNA (shRNA) mediated contactin-1 (CNTN1) gene silencing on growth of human breast cancer cell line MDA-MB-468 transplanted tumors in nude mice.MethodsEighteen nude mice (4-week-old male BALB/c) were randomly equally divided into three groups: blank control group, empty vector group, and silencing group. The MDA-MB-468 cells (blank control group), MDA-MB-468 cells transfected by nonsense shRNA (empty vector group), and MDA-MB-468 cells transfected by shRNA (silencing group) were collected in the logarithmic growth period, respectively. The subcutaneous tumor models of nude mice were prepared by the subcutaneous injection of the different group cells. The tumor growth was observed and the expressions of CNTN1 and Ki-67 proteins in the transplanted tumor were detected by the immunohistochemistry.ResultsThe xenograft models of human breast cancer cells were established successfully. The tumor growth in the silencing group was significantly slower than that of the other two groups at every 3 d point (P<0.05). The tumor volume and the tumor weight in the silencing group were significantly smaller or slighter than those of the other two groups at day 18 (P<0.05). The positive rates of CNTN1 and Ki-67 protein expressions in the tumor tissues of the silencing group were lower than those of the other two groups (P<0.05), respectively.ConclusionSilencing expression of CNTN1 gene might inhibit growth of breast cancer cell line MDA-MB-468 transplanted tumors in mude mice.