HCT116 VIM RFP
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The HCT116 VIM RFP reporter cell line, derived from the parental HCT116 (ATCC CCL-247™) colorectal carcinoma cell line, was created using CRISPR/Cas9 gene editing technology. The HCT116 VIM RFP reporter cell line carries a knock-in red fluorescent protein (RFP) reporter, which was integrated before the stop codon at the last exon of the endogenous vimentin (VIM) gene.
Epithelial to mesenchymal transition (EMT) has been recognized to play an important role in cancer cell metastasis and drug resistance. The EMT pathway is of increasing interest as a novel therapeutic avenue in the treatment of cancer. HCT116 VIM RFP cell line (ATCC CCL-247EMT™) was created using the CRISPR-Cas9 platform, in which a red fluorescent protein (RFP) reporter was integrated before the stop codon at the last exon of the endogenous vimentin gene, a widely used mesenchymal cell marker. The integrity of the VIM RFP knock-in allele has been verified at genomic, transcriptional, and translational levels. In HCT116 VIM RFP cells, vimentin-RFP expression can be robustly turned on in response to miR-200 family inhibitor treatment, as previously reported (Park et al, Genes Dev, 22: 894-907, 2008). In contrast, the expression of E-cadherin, a marker of epithelial cells, is significantly reduced upon miR-200 inhibitor treatment. In addition, hypomethylating agent 5-Aza-2’-Deoxycytidine treatment can induce an increase in VIM RFP expression effectively. The HCT116 VIM RFP cell line could be a useful in vitro cell model for dissecting the molecular switches underlying EMT and for identifying compounds that target EMT in colorectal cancer.
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
The STR profile of the HCT-116 VIM RFP reporter cell line is approximately an 80% match to the parental HCT-116 cell line indicating that the cell lines are related (derived from common ancestry). The STR differences are likely attributed to microsatellite instability in the HT-116 cell line, which is a common feature of this cell line.1,2,3
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Park SM, et al. The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev 22(7): 894-907, 2008. PubMed: 18381893
Schroy PC, et al. Detection of p21ras mutations in colorectal adenomas and carcinomas by enzyme-linked immunosorbent assay. Cancer 76: 201-209, 1995. PubMed: 8625092
Brattain MG, et al. Heterogeneity of malignant cells from a human colonic carcinoma. Cancer Res. 41: 1751-1756, 1981. PubMed: 7214343
Sun L, et al. Autocrine transforming growth factor-beta 1 and beta 2 expression is increased by cell crowding and quiescence in colon carcinoma cells. Exp. Cell Res. 214: 215-224, 1994. PubMed: 8082724
Santoro IM, Groden J. Alternative splicing of the APC gene and its association with terminal differentiation. Cancer Res. 57: 488-494, 1997. PubMed: 9012479