scholarly journals Evaluation of Thymidylate Synthase Protein Expression by Western Blotting and Immunohistochemistry on Human Colon Carcinoma Xenografts in Nude Mice

2002 ◽  
Vol 50 (12) ◽  
pp. 1633-1640 ◽  
Author(s):  
Massimo Derenzini ◽  
Lorenzo Montanaro ◽  
Alessandra Chillà ◽  
Elena Tosti ◽  
Claudio Ceccarelli ◽  
...  
Keyword(s):  
Growth Rate ◽  
Cell Growth ◽  
Protein Expression ◽  
Western Blotting ◽  
Thymidylate Synthase ◽  
Human Colon ◽  
Growth Fraction ◽  
Tumor Cell Growth ◽  
Mass Growth ◽  
Tumor Mass

In this study we investigated the relationship between thymidylate synthase (TS) protein expression, evaluated by Western blotting analysis and by immunohistochemistry (IHC), and growth rate in human colon xenograft tumors in nude mice. Human colon cancer cell lines were used to induce xenograft tumors and the tumor mass growth rate was calculated by measuring tumor size variations over time. TS 106 monoclonal antibody was used for both Western blotting and IHC TS detection. Tumor cell growth fraction was measured by Ki67/MIB1 immunolabeling and tumor cell growth rate by evaluating the mean nucleolar size in silver-stained sections. TS Western blotting values were related to tumor mass growth rate ( p<0.001) and cell growth rate ( p=0.002) but not to cell growth fraction ( p=0.676). The degree of the IHC staining showed only a trend to be associated with TS protein expression measured on Western blotting, and was not related either to tumor mass growth or cell proliferation rate. Tumor xenografts were also characterized for TS promoter tandem repeat and p53 status. No relationship was observed between these variables and TS expression evaluated by both Western blotting and IHC analysis. Our results demonstrate that TS expression evaluated by Western blotting analysis is directly related to the tumor mass growth rate and question the use of the IHC approach to obtain precise quantitative information on TS expression in tumor samples.

Let-7a inhibits tumor cell growth and metastasis by directly targeting RTKN in human colon cancer

2016 ◽  
Vol 478 (2) ◽  
pp. 739-745 ◽  
Author(s):  
Bin Li ◽  
Peng Chen ◽  
Yanxiang Chang ◽  
Jingpeng Qi ◽  
Hui Fu ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cell Growth ◽  
Tumor Cell ◽  
Human Colon ◽  
Tumor Cell Growth ◽  
Human Colon Cancer

scholarly journals Polymorphism of thymidylate synthase gene associated with its protein expression in human colon cancer

2008 ◽  
Vol 14 (4) ◽  
pp. 617 ◽  
Author(s):  
Kai-Huan Yu ◽  
Wei-Xing Wang ◽  
You-Ming Ding ◽  
Hui Li ◽  
Ze-Sheng Wang
Keyword(s):  
Colon Cancer ◽  
Protein Expression ◽  
Thymidylate Synthase ◽  
Human Colon ◽  
Human Colon Cancer

Preliminary Functional Studies of hnRNPK in Progression of CML

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4229-4229
Author(s):  
Liu Xiaoli ◽  
Hongqian Zhu ◽  
Song Zhang ◽  
Qingfeng Du ◽  
Junmei Gong ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Growth ◽  
Protein Expression ◽  
Western Blotting ◽  
Molecular Mechanisms ◽  
Mononuclear Cells ◽  
Blast Crisis ◽  
Terminal Phase ◽  
Reasonable Assumption ◽  
Hematopoietic Stem

Abstract The blast crisis(BC) is terminal phase of chronic myeloid leukemia (CML), which is accompanied by an increase in both BCR/ABL mRNA and protein level and imatinib(IM) resistance. The BCR/ABL oncoprotein is responsible for inducing and sustaining the leukemic phenotype through its deregulated tyrosine kinase activity which is essential for recruitment and induction of signaling pathways leading to cytokine-independent proliferation, resistance to apoptosis, and impaired differentiation of BCR/ABL-expressing myeloid and lymphoid progenitors. However, the molecular mechanisms responsible for blastic transformation remain poorly understood, although a reasonable assumption is that the unrestrained activity of BCR/ABL in hematopoietic stem/progenitor cells is the primary determinant of disease progression. We had analyzed the changes of protein expression between the bone marrow mononuclear cells in CML-CP and that in CML-BC by the technique of proteome. Compared to CML-CP, the protein expression of heterogeneous nuclear ribonucleoprotein K(hnRNPK) increases in CML-BC. In present study, the over-expression of hnRNPK in CML-BC was verified by western blotting. Furthermore, the expression of hnRNPK increases in a new imatinib-resistant BCR/ABL-positive cell line, K562-R, than in the wild-type K562. After K562 and K562-R were treated by different inhibitors such as PD98059, LY294002, AG490 and imatinib, the protein expression and mRNA level of hnRNPK were detected by western blotting and QRT-PCR. Inhibition of BCR-ABL by imatinib significantly decreased hnRNPK expression in K562 and inhibition of MEK by PD98059, downstream of the BCR-ABL signaling cascade, decreased hnRNPK expression in both K562 and K562-R cells. In addition, down-regulation of hnRNPK by small double-stranded RNA specifically complementary to hnRNPK (siRNA-hnRNPK) can inhibite cell growth and induce maximal G2/M block at 48 hours in both K562 and K562-R cells, whereas cell appototis was observed only at 72 hours by AnnexinV-PI assay. The combination of siRNA-hnRNPK and imatinib induce more cell death in K562-R cells than either treatment alone. Our data therefore suggest that hnRNPK is regulated by the BCR-ABL/MAPK cascade in Ph+ CML. The results that down-regulating hnRNPK expression induced cell-growth arrest and subsequent cell death suggest the potential therapeutic utility of this strategy in patients with CML.

scholarly journals A “combination oligonucleotide” antisense strategy to downregulate thymidylate synthase and decrease tumor cell growth and drug resistance

2003 ◽  
Vol 10 (4) ◽  
pp. 278-286 ◽  
Author(s):  
Randal W Berg ◽  
Peter J Ferguson ◽  
Mark D Vincent ◽  
D James Koropatnick
Keyword(s):  
Drug Resistance ◽  
Cell Growth ◽  
Tumor Cell ◽  
Thymidylate Synthase ◽  
Tumor Cell Growth

FAPP2 promotes tumor cell growth in human colon cancer through activation of Wnt signaling

2019 ◽  
Vol 374 (1) ◽  
pp. 12-18 ◽  
Author(s):  
Jingde Chen ◽  
Li Li ◽  
Zhuqing Zhou ◽  
Shijun Yu ◽  
Yandong Li ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cell Growth ◽  
Tumor Cell ◽  
Wnt Signaling ◽  
Human Colon ◽  
Tumor Cell Growth ◽  
Human Colon Cancer

Effects of PHA-665752 and Cetuximab Combination Treatment on In Vitro and Murine Xenograft Growth of Human Colorectal Cancer Cells with KRAS or BRAF Mutations

2018 ◽  
Vol 18 (3) ◽  
pp. 278-286 ◽  
Author(s):  
Yi-Tao Jia ◽  
Dong-hai Yang ◽  
Zhaolong Zhao ◽  
Xiao-Hui Bi ◽  
Wei-Hua Han ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Growth ◽  
Protein Expression ◽  
Cell Lines ◽  
Western Blotting ◽  
Combination Treatment ◽  
Human Colorectal Cancer ◽  
Hct 116 ◽  
Ht 29

Background: It remains unknown whether blockade of c-Met signaling and epidermal growth factor receptor signaling is effective in suppressing the growth of human colorectal cancer (CRC) cells. In this study, we investigated the effects of the c-Met inhibitor PHA-665752 alone and in combination with cetuximab on the growth of human CRC cells in vitro and in mouse xenografts. Methods: Human CRC cell lines (Caco2, HCT-116, and HT-29) and mice bearing HCT-116 xenografts were treated with cetuximab in the absence or presence of PHA-665752. Cell viability and apoptosis were examined using the MTT and TUNEL assays, respectively. Vimentin was measured by immunohistochemistry as a marker for epithelial-to-mesenchymal transition. Western blotting was used to determine signaling protein expression levels. Results: The MTT assay showed that the growth of Caco2, HCT-116, and HT-29 cells was inhibited by PHA-665752 in a dose-dependent manner, but only Caco2 cell growth was suppressed by cetuximab. Combination treatment with PHA-665752 and cetuximab inhibited the proliferation of Caco2 cells and RAS mutant CRC cell lines. However, relative to the PHA-665752-alone treatment group, HT-29 cells with a BRAF mutation showed no noticeable effect. The mean tumor volume in mice treated with cetuximab in combination with PHA-665752 was significantly smaller than that in the mice treated with only cetuximab (P = 0.033) or PHA-665752 (P < 0.01). Similarly, the expression of vimentin in the mice treated with PHA-665752 in combination with cetuximab was significantly lower than that in the mice treated with cetuximab or PHA-665752 alone (P < 0.05 in each case). TUNEL assays revealed that treatment with PHA-665752 in combination with cetuximab markedly increased CRC cell apoptosis. Western blotting analysis of signaling protein expression showed that PHA- 665752 inhibited Met phosphorylation (P < 0.05). In addition, treatment with cetuximab alone or in combination with PHA-665752 effectively inhibited EGFR phosphorylation (P < 0.05). Conclusion: Combination treatment with PHA-665752 and cetuximab suppressed in vitro and in vivo CRC cell growth more than treatment with either agent alone did.

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