ABSTRACT
Tumor cell line culture technologies emerged as a strong impetus to the development of tumor biology research.However,cell line tumor model presented limited predictive value and instable properties in the clinical translational research of new cancer therapies.Recently Patient-derived Xenograft Model (PDX) has aroused researcher's attention and been widely applied to new cancer therapy research.This new model could highly retain the stroma heterogeneity、histological characteristics and molecular diversity of the original tumor,and is expected to bring new breakthroughs in individualized treatment of tumor patients.
ABSTRACT
Objective To explore the expression of V-ATPase in colon cancer and its clinical significance. Methods Detecting the expression of V-ATPase mRNA in 20 paired of colon tumor tissues and normal tissues by using reverse transcription-polymerase chain reaction ( RT-PCR) and real-time quantitative polymerase chain reaction( Real-time PCR) , and testing the expression of V-ATPase protein by immu-nohistochemistry of EnVinsion. Results The expression of V-ATPase mRNA in tumor tissues and its paired normal tissues were (5. 37 ± 0. 44) and (2. 03 ± 0. 35)(P<0. 01). The positive immunohistochemistry of V-ATPase in tumor tissues and its paired normal tissues were 69. 1%(47/68) and 5. 8%(4/68) respectively, and the positive expression were primarily in cytoplasm and cytomembrane. Overexpression of V-ATPase was associated with tumor stage (P<0. 05), lymph node metastasis (P=0. 044), distant metastasis (P=0. 049), vessel in-vasion (P=0. 044) and differentiation (P<0. 001). Conclusion Overexpression of V-ATPase plays a significant role in the carcinogene-sis and the progression of colon cancer, which might be an important postoperative therapeutic target.
ABSTRACT
Objective To evaluate the effect of sustained silencing Forkhead box Ml (F0XM1) gene by short-hairpin RNA (shRNA) expression vector on cell growth of hepatocelluar carcinoma (HCC) in vitro.Methods Four shRNA expression vectors targeting different sequences of human F0XM1 mRNA were constructed.The expression vector with the best interfering effect and the negative control plasmid were used to transfect HCC cell line QGY-7703, stably transfected cell clones were selected by neomycin (G418).Cell growth was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation was assessed by clonogenic assay.Cell apoptosis was detected by double staining with APC conjugated Annexin V and PI.Results F0XM1 protein was detected with different levels in all these studied human cell lines.The expression vector shRNA-1026 exhibited excellent interference effect after transient transfection, which showed 38.5% and 53.2% reduction of FOXM1 mRNA and protein level respectively.The growth of QGY-7703 cells was inhibited after stable inhibition of FOXM1 expression by shRNA-1026, which was indicated by decreased absorbance value of the test group after culture for 48, 72 and 96 h compared to control group (t = 10.830,3.578 and 5.734 respectively, P ＜ 0.05).Stable inhibition of F0XM1 also led to reduced colony formation ( t = 5.336, P ＜ 0.05 ) and increased apoptosis of QGY-7703 cells in comparison to control cells (t = 6.827, P ＜ 0.05 ).Conclusions Stable silencing F0XM1 gene by shRNA suppresses the growth of HCC cells in vitro.