Translation of an atypical human cDNA requires fidelity of apurine-pyrimidine repeat region and recoding

Petty, Aaron; Dick, Charles L.; Lindsey, J. Suzanne

doi:10.1016/j.gene.2008.02.006

Cited by 3 publications

(6 citation statements)

References 33 publications

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“…The effect of amino terminus FLAG-tagged Mig-7 (FLAGMig-7) overexpression in stably transfected HT29 colon carcinoma cells was quantitatively analyzed compared to vector alone transfected cells. We chose this cell line because it does not endogenously express Mig-7 and it stably overexpresses FLAGMig-7 at the protein level (2;3;20). Overexpression in HT29 cells significantly increased the total number of invaded cells by 8-fold (p = 0.002) over control (Figure 1a).…”

Section: Resultsmentioning

confidence: 99%

“…the frame that did not produce protein (20), were used to stimulate MC in vitro . In addition, these peptides included overlapping sequences (see Methods) to prevent the possible omission of an epitope.…”

Section: Resultsmentioning

confidence: 99%

“…These data are not surprising because even with 40 cycles of PCR Mig-7 mRNA is not detected in 25 different normal human tissues (n=6 each tissue) or in blood from normal subjects (2;4). This specificity of Mig-7 expression may be due to multiple tumor microenvironment factors such as ligation of αvβ5 integrin and multiple growth factors leading to Mig-7 transcription (2;3) as well as multiple events required for Mig-7 translation (20). The percentage of total breast carcinoma tissues staining for Mig-7 was 53%.…”

Section: Discussionmentioning

confidence: 99%

“…Data from the present studies show that inhibiting Mig-7 function by antibody treatment or expression by Mig-7 shRNA leads to decreased levels of activated MT1-MMP, suggesting a mechanism by which Mig-7 promotes the above cellular events. MT1-MMP can be activated through the “cysteine switch” mechanism (29;30) and Mig-7 is highly cysteine-rich (2;20). Additional studies beyond the scope of this work are needed to determine if one or more Mig-7 cysteine residues play a role in MT1-MMP activation.…”

Section: Discussionmentioning

confidence: 99%

“…Methods for constructing expression vectors, FLAGMig-7 and shRNA, as well as transfecting, selecting and culturing HT29 colon carcinoma, HEC1A, RL95 endometrial carcinoma (2;3;20), and MCF-7 breast carcinoma (21) cell lines were previously described. Mig-7 sequence (Accession DQ080207) of the previously unpublished shRNA construct insert, 4-2 antisense -loop- sense, is TCATTCACCTGCTATAGAC TTCAAGAGA GTCTATAGCAGG-TGAATGA (bp 1303 to 1321).…”

Section: Methodsmentioning

confidence: 99%

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Targeting Migration inducting gene-7 inhibits carcinoma cell invasion, early primary tumor growth, and stimulates monocyte oncolytic activity

Petty

Wright

Yenderrozos

et al. 2009

Molecular Cancer Therapeutics

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Expression of Migration inducting gene-7 (Mig-7) is limited to tumor cells and to date not found in normal tissues. Multiple tumor microenvironment factors, such as epidermal and hepatocyte growth factors, in concert with αvβ5 integrin ligation, induce Mig-7 mRNA expression. Gain or loss of Mig-7 protein studies shows that Mig-7 promotes invasion of colon and endometrial carcinoma cells. These data led us to hypothesize that targeting Mig-7 through various methods could decrease invasion, enhance monocyte cell killing of tumor cells, and inhibit disease progression. To begin testing this hypothesis, an in vitro chemoinvasion assay of endometrial carcinoma cells treated with Mig-7-specific or control antibodies was used. Mig-7 antibody significantly reduced invasion by >60% compared with controls. In another approach to test this hypothesis, an in vitro analysis of peptidestimulated human peripheral blood monocyte cells and their killing of MCF-7 breast carcinoma cells was used. Mig-7 peptide treatment increased monocyte cell tumor necrosis factor expression and killing of MCF-7 cells 30-fold over no peptide stimulation and 3-fold over MUC-1 or control peptide treatments. Furthermore, stably expressing Mig-7-specific short hairpin RNA resulted in significantly reduced Mig-7 protein levels and early primary tumor growth in a xenograft nude mouse model. Reduced phosphorylation of ERK1/2, Akt, and S6 kinase as well as decreased membrane-type 1 matrix metalloproteinase activity were mechanisms through which Mig-7 protein caused these effects. Based on these collective data, Mig-7 expression could be a potential candidate for future targeted cancer therapies.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Targeting Migration inducting gene-7 inhibits carcinoma cell invasion, early primary tumor growth, and stimulates monocyte oncolytic activity

Petty

Wright

Yenderrozos

et al. 2009

Molecular Cancer Therapeutics

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Isoforms of wild type proteins often appear as low molecular weight bands on SDS‐PAGE

Zhang

Lou

Shen

et al. 2014

Biotechnology Journal

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Immunoblotting, after polyacrylamide gel electrophoresis with sodium dodecyl sulfate (SDS-PAGE), is a technique commonly used to detect specific proteins. SDS-PAGE often results in the visualization of protein band(s) in addition to the one expected based on the theoretical molecular mass (TMM) of the protein of interest. To determine the likelihood of additional band(s) being nonspecific, we used liquid chromatography - mass spectrometry to identify proteins that were extracted from bands with the apparent molecular mass (MM) of 40 and 26 kD, originating from protein extracts derived from non-malignant HEK293 and cancerous MDA-MB231 (MB231) cells separated using SDS-PAGE. In total, approximately 57% and 21% of the MS/MS spectra were annotated as peptides in the two cell samples, respectively. Moreover, approximately 24% and 36.2% of the identified proteins from HEK293 and MB231 cells matched their TMMs. Of the identified proteins, 8% from HEK293 and 26% from MB231 had apparent MMs that were larger than predicted, and 67% from HEK293 and 37% from MB231 exhibited smaller MM values than predicted. These revelations suggest that interpretation of the positive bands of immunoblots should be conducted with caution. This study also shows that protein identification performed by mass spectrometry on bands excised from SDS-PAGE gels could make valuable contributions to the identification of cancer biomarkers, and to cancer-therapy studies.

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To Know How a Gene Works, We Need to Redefine It First but then, More Importantly, to Let the Cell Itself Decide How to Transcribe and Process Its RNAs

Jia¹,

Chen²,

Ma³

et al. 2015

Int. J. Biol. Sci.

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Recent genomic and ribonomic research reveals that our genome produces a stupendous amount of non-coding RNAs (ncRNAs), including antisense RNAs, and that many genes contain other gene(s) in their introns. Since ncRNAs either regulate the transcription, translation or stability of mRNAs or directly exert cellular functions, they should be regarded as the fourth category of RNAs, after ribosomal, messenger and transfer RNAs. These and other research advances challenge the current concept of gene and raise a question as to how we should redefine gene. We can either consider each tiny part of the classically-defined gene, such as each mRNA variant, as a “gene”, or, alternatively and oppositely, regard a whole genomic locus as a “gene” that may contain intron-embedded genes and produce different types of RNAs and proteins. Each of the two ways to redefine gene not only has its strengths and weaknesses but also has its particular concern on the methodology for the determination of the gene's function: Ectopic expression of complementary DNA (cDNA) in cells has in the past decades provided us with great deal of detail about the functions of individual mRNA variants, and will make the data less conflicting with each other if just a small part of a classically-defined gene is considered as a “gene”. On the other hand, genomic DNA (gDNA) will better help us in understanding the collective function of a genomic locus. In our opinion, we need to be more cautious in the use of cDNA and in the explanation of data resulting from cDNA, and, instead, should make delivery of gDNA into cells routine in determination of genes' functions, although this demands some technology renovation.

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Translation of an atypical human cDNA requires fidelity of apurine-pyrimidine repeat region and recoding

Cited by 3 publications

References 33 publications

Targeting Migration inducting gene-7 inhibits carcinoma cell invasion, early primary tumor growth, and stimulates monocyte oncolytic activity

Targeting Migration inducting gene-7 inhibits carcinoma cell invasion, early primary tumor growth, and stimulates monocyte oncolytic activity

Isoforms of wild type proteins often appear as low molecular weight bands on SDS‐PAGE

To Know How a Gene Works, We Need to Redefine It First but then, More Importantly, to Let the Cell Itself Decide How to Transcribe and Process Its RNAs

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