Killing Cancer: The Effect of Galectin-7 Overexpression in Oral Cancer Cell Line

Overview

Objectives/Goals To examine the effect of galectin-7 overexpression in oral cancer cell line. Aim 1. Clone galectin-7/pcDNA3.1 expression vector Aim 2. Transfect into galectin-7/pcDNA3.1 or pcDNA3.1 into SCC4 and SCC9 Aim 3. Serum-starvation and assay for apoptosis (examine morphology, nuclear condensation and DNA fragmentation) Methods/Materials Our study has examined our hypothesis that exogenous expression of galectin-7 in oral cancer cell line will facilitate apoptosis. We tested this by constructing galectin-7 expression vector in pcDNA3.1 and transfecting it into SCC4 and SCC9 oral squamous cell carcinoma cell line. A 550-bp restriction fragment containing full-length galectin-7 was ligated into pcDNA3.1 mammalian expression vector. The recombinant construct was transformed into Escherichia coli DH5a strain and ampicillin resistant colonies were selected. Gal-7/pcDNA3.1 was digested with HindIII/BamHI and the restriction fragment fractionated on a 1% agarose gel. Transient transfection of gal-7/pcDNA3.1 resulted in expression of 700 bp gal-7 mRNA in SCC9 cells. Under serum starvation exogenous expression of gal-7 in SCC9 resulted in apoptosis sensitivity compared to cells with or without pcDNA3.1 vector. Results Plasmid isolation and subsequent restriction digest showed that some of the ampicillin resistant colonies contained the galectin-7/pcDNA3.1 construct. Analysis of transiently transfected cells showed that galectin-7 mRNA and galectin-7 protein were expressed in SCC4 and SCC9 that was transfected with galectin-7/pcDNA3.1. Serum starvation of SCC4 and SCC9 overexpressing galectin-7 resulted in cells with increased membrane blebbing along with cytosolic shrinkage. Conclusions/Discussion We conclude that overexpression of galectin-7 in oral cancer cells resulted in sensitivity to apoptosis. We plan to: ·Obtain results from cells transfected with pcDNA3.1 control that did not express neither mRNA or protein for galectin-7 ·Overexpress Galectin-7 in HPV16-Immortalized Human Oral Keratinocytes

Competition history

• CSEF 2004 Biochemistry/ Molecular Biology · Entry S0408

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