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1.
Chinese Journal of Cancer Biotherapy ; (6): 590-597, 2021.
Article in Chinese | WPRIM | ID: wpr-882197

ABSTRACT

@#[摘 要] 目的:体内外实验探讨木鳖子单体化合物对羟基桂皮醛[Momordica cochinchinensis(Lour.)Spreng.p-hydroxycinnamaldehyde,CMSP]对小鼠黑色素瘤移植瘤生长和转移的影响及其作用机制。方法:建立荷瘤小鼠动物模型,并将18只C57BL/6小鼠随机分成3组(每组6只):对照组(腹腔注射0.1 ml生理盐水)、CMSP治疗组(分别腹腔注射0.1 ml 1、2 mg/ml CMSP),给药的第5天开始,每次给药前用卡尺分别测量和计算小鼠移植瘤的体积,实验结束后称量移植瘤的质量;H-E染色后光镜观察肝组织的病理学变化;免疫组织化学SP法观察移植瘤组织E-cadherin和vimentin蛋白的表达。采用细胞划痕和Transwell实验分别检测CMSP实验组(10、20 µg/ml)黑色素瘤B16细胞24、48 h的迁移能力,qPCR法检测CMSP处理24 h后B16细胞EMT相关mRNA表达,WB法检测CMSP处理B16细胞48 h后β-catenin、p-β-catenin(Ser675)、vimentin和E-cadherin蛋白的表达水平。结果:CMSP治疗组小鼠移植瘤平均体积和肿瘤质量明显降低(均P<0.05);对照组小鼠肝脏中转移灶的数量明显多于CMSP(1、2 mg/kg)治疗组(均P<0.05),CMSP(2 mg/kg)处理组小鼠的肝组织内未发现明显转移灶。CMSP治疗组(1、2 mg/kg)移植瘤组织中E-cadherin蛋白表达水平明显高于对照组(均P<0.05),而vimentin蛋白表达显著低于对照组(均P<0.01)。体外实验中,CMSP实验组(10、20 μg/ml)B16细胞24、48 h后划痕愈合率较对照组均明显降低(均P<0.05)。20 μg/ml CMSP处理B16细胞24、48 h后穿过Transwell小室的细胞数较对照组则显著下降(均P<0.01)。CMSP(10、20 μg/ml)处理B16细胞后β-catenin mRNA表达水平较对照组明显降低(均P<0.01),E-cadherin mRNA表达水平则明显升高(均P<0.05),而vimentin mRNA表达水平在10 μg/ml处理组与对照组相比差异无统计学意义(P>0.05),20 μg/ml处理组则明显降低(P<0.01)。与对照组相比,CMSP实验组(10、20 μg/ml)处理B16细胞后β-catenin、p-β-catenin和vimentin蛋白表达均显著降低(均P<0.01),而E-cadherin蛋白表达则明显升高(均P<0.01)。结论:CMSP能够抑制小鼠黑色素瘤移植瘤的生长和转移,其作用机制可能与抑制wnt/β-catenin通路的活性相关。

2.
Chinese Journal of Cancer Biotherapy ; (6): 1018-1023, 2020.
Article in Chinese | WPRIM | ID: wpr-825754

ABSTRACT

@#[Abstract] Objective: To observe the effect of allogeneic platelets transfusion on the invasion and metastasis of human lung cancer A549 cells, and to preliminarily explore its mechanism of action. Methods: Eighty-nine patients with advanced lung cancer, who had received platelet transfusion in the Chemotherapy Department of Fourth Hospital of Hebei Medical University between January 2017 and December 2018, were enrolled in this study. The study cells were randomized into Ctrl group (A549 cells co-incubated with culture medium), Before group, and After group (A549 cells co-incubated with plasma Before and After platelet transfusion, respectively). The migration and invasion of A549 cells co-cultured with plasma before and after platelet transfection were detected by Scratch and Transwell experiments. The expression of MMPs, TIMPs and epithelial-mesenchymal transition (EMT) related proteins E-cadherin, N-cadherin and Vimentin, as well as vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2) were detected by Western blotting (WB) method. Results: The scratch healing ability of A549 cells in After group was significantly higher than that of Ctrl group and Before group [(73.67±2.60)% vs (58.33±2.33)%, (35.33±2.03) %; P<0.01, vs Ctrl group; P<0.05, vs Before group], and there was also a significant difference between Before group and Ctrl group (P<0.05). The results of cell migration experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(69.67±7.84) vs (18±2.08) and (39.33±2.03), all P<0.01]. The cell invasion experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(59.34±3.46) vs (18.34±1.56) and (37.58±2.79), all P<0.01]. When A549 cells were co-incubated with plasma before and after platelet transfusion for 48 h, it was found that the expressions of MMP9 and MMP2 were increased (P<0.05), while their inhibitors TIMP1 and TIMP2 were decreased (P<0.01); the expressions of EMT-related proteins N-cadherin and Vimentin were increased (P<0.05), but E-cadherin was decreased (P<0.01); the expressions of angiogenesis related proteins VEGF and VEGFR2 were increased (P<0.05). Conclusion: Alloplatelets transfusion can promote the invasion and metastasis of lung cancer A549 cells, which may be realized by regulation of the expressions of EMT, metallomatrix protease and vascular growth factor-related proteins.

3.
Electron. j. biotechnol ; 35: 10-17, sept. 2018. tab, graf
Article in English | LILACS | ID: biblio-1047827

ABSTRACT

Alanine mother liquor, a type of industrial waste from alanine fermentation, was used as a nitrogen source to produce docosahexaenoic acid (DHA) by Schizochytrium sp. B4D1. The results indicated that yeast extract could trigger the utilization of the alanine mother liquor. Additionally, the alanine can be quenched during the culture, which aids in DHA accumulation. The medium components were optimized via response surface methodology as follows: 99.98-g/L glucose, 0.05-g/L yeast extract and a 183.17 dilution factor of the alanine mother liquid (v/v, with an alanine content of 0.72 g/L) and 17.98% inoculum concentration (v/v). Finally, in a 50-mL shake-flask fermentation, the DHA yield was 2.29 g/L.


Subject(s)
Docosahexaenoic Acids/biosynthesis , Alanine/metabolism , Stramenopiles/metabolism , Yeasts , Intercellular Signaling Peptides and Proteins/isolation & purification , Alanine/analysis , Fermentation , Glucose , Industrial Waste
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