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BACKGROUND: Acellular scaffolds have been widely used in the study of various tumors in recent years. The spatial arrangement, biomechanical properties and biocompatibility of the scaffolds are helpful to restore the microenvironment of tumor cell growth. OBJECTIVE: To investigate the characteristics and advantages of porcine colon acellular scaffold as an in vitro model of colon cancer. METHODS: Acellular scaffolds of porcine colon were prepared by soaking fresh porcine colon with 2% SDS, 1% TritonX-100 and 0.5% EDTA combined with repeated shaking. Human colon cancer HCT116 cells were inoculated on the mucous surface of porcine colon acellular scaffold. The growth of cells on the scaffold was observed by live-death staining. The morphology of cells on the scaffold was observed by phalloidin staining, and the longitudinal growth of cells on the scaffold was observed by hematoxylin-eosin staining and immunofluorescence staining. RESULTS AND CONCLUSION: (1) Live-death staining showed that on the first day of culture, HCT116 cells could well gather in the pores of the mucosal layer of the scaffolds, and few dead cells were found. On the third day, the cells gradually spread out of the pores, and a small number of cells grew and connected into pieces, and there were no dead cells. On the 7th day, the cell growth density further increased and grew into a sheet on the surface of the mucous layer, and there were no dead cells and exfoliated cells. (2) On the 7th day of culture, hematoxylin-eosin staining and immunofluorescence staining showed that some HCT116 cells could grow into clusters in the pores, and some cells continued to grow to the submucosa along the pores, showing the characteristics of invasive growth of intestinal cancer in situ. (3) On the 7th day of culture, the phalloidin staining showed that HCT116 cells were in close contact with the scaffold mucosa at the ultrastructural level, and had well-differentiated epithelial morphological characteristics. (4) The result shows that porcine colon acellular scaffold can be used as a carrier for three-dimensional culture of colon cancer cells.
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ObjectiveTo explore the role of miR-34a in the proliferation, migration and invasion of hepatocellular carcinoma (HCC) cells.MethodsHepG2 and Huh-7 cells were transfected with miR-34a constructs by using lipofection. qPCR was applied to detect the expression of miR-34a. Cell proliferation assay, colony formation assay, cell migration and invasion assay were used to elucidate the invasive growth of transfected and un-transfected cells. The subcutaneous xenograft in nude mice was used to verified the anti-tumor effect of miR-34a in vivo. TargetScan, miR-WALK and luciferase assay were performed to explore the target of miR-34a in HCC.ResultsCompared with the overexpression control group and blank group, the expression of miR-34a in the miR-34a overexpression group cells were (2.727±0.173) and (4.042±0.104), respectively (P<0.05). The levels of miR-34a in overexpression control and blank groups were (0.003±0.030) and (0.005±0.027) (P＞0.05). Compared with the overexpression control groups, cell proliferation, colony formation, migration and invasion of HCC cells overexpressing miR34a were all impeded with statistically significant differences (P<0.05). In vivo, tumor weight and tumor volume were significantly reduced in the miR-34a treatment group. The results of luciferase reporter gene indicated that c-Met was the direct target of miR-34a in HCC.ConclusionsUp-regulation of miR-34a in hepatocellular carcinoma can inhibit the proliferation, migration and invasion of HCC cells, and its repression may be benefited by declining c-Met signaling in HCC.
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Alveolar echinococcosis (AE) is a potentially fatal zoonosis caused by Echinococcus multilocularis (E .m) , primary located in the liver and showed malignant invasion .AE can develop to the lungs and brain metastases .Operation treat-ment and medication therapeutic efficacy show poor effect .The mechanism of AE invasive growth and metastasis is unclear . Angiogenesis is a highly regulated behavior in normal tissues .Abnormal angiogenesis is one of the key signs of malignant tumor ,and VEGF is one of the key mediators of angiogenesis .Numerous studies have shown that VEGF and CD34 plays an important role in the formation of tumor angiogenesis and invasive growth process .This article discussed the role of angiogene-sis in AE aggressive ,aimed to provide a new way for the clinic conservative treatment for patients with advanced stage ,preven-ting recurrence after operation ,and adjunctive therapy of operation and drug .
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Objegtive To investigate the effect and significance of cinobufagin on the expression of CD44s in ovarian carcinoma 3AO cells cultured in vitro.Methods Ovarian carcinoma 3AO cells were cultured in vitro;after the intervention of tinobufagin with different concentration,the expression of CD44s was measured by RT-PCR,and the cellular apoptosis was tested by flow cytometry.Results Ovarian carcinoma 3AO cells expressed CD44S mRNA of 1.3±0.1 and the cellular apoptofic rate was(2.31±0.98)%:0.25 mg/ml cinobufagin did not affect CD44s mRNA and cellular apoptofic rate(P ＞0.05):when 2.5 mg/ml cinobufagin was intervened,the expression of CD44s mRNA was 1.0±0.1 and the cellular apoptotic rate Was(28.69±4.16)%,showing significant difference comparing with the control group of ovarian carcinoma 3AO ceils(P＜0.05):the intervention of 25 mg/ml cinobufagin was similar to that of 0.25 mg/ml cinobuhgin(P＞0.05).3AO cells.
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PURPOSE: To report a patient with malignant conjunctival melanoma with extensive corneal invasive growth. METHODS: A 63-year-old woman with 9 x 7 mm sized dark brown colored conjunctival mass with extensive corneal invasive growth over a 3-month history was seen. The mass was removed and biopsy was done. Fontana-Masson, S-100 protein, and HMB stainings were used to confirm the diagnosis of malignant melanoma. RESULTS: The tumor was easily separated from the corneal surface and adjacent conjunctiva was excised. Fontana-Masson, S-100 protein, and HMB stainings revealed corneally-displaced malignant conjunctival melanoma. There was no evidence of metastasis. CONCLUSIONS: Corneally displaced malignant conjunctival melanoma was very rare. A good surgical result was obtained with its removal.