A review of the article by Jankowska, et al., 2008, titled “Reduction of human chorionic gonadotropin beta subunit expression by modified U1 snRNA caused apoptosis in cervical cancer cells”
Jankowska, et al in their article published in the journal “Molecular Cancer” demonstrate the expression of hCG-beta in cervical cancer and examine the functional significance of hCG- in-vitro cervical cancer cells by making use of modified U1snRNA to silence the mRNA of hCG-. Silencing of the maturation of mRNA by use of a novel silencing technique results in inhibition of growth and apoptosis of cervical cancer cells.
Lipofectamine 2000 and Fu-GENE HD transfection reagents were utilized for transfection of the anti-hCG- snRNA into the cervical carcinoma (Hela) cells. Lipofectamine 2000, which is a cationic liposome based transfection reagent forms complex with the negatively charged RNA (snRNA) and DNA molecules and allows their passage through negatively charged plasma membrane of the cells (Dalby et al., 2004). The authors report to have observed a 30% transfection efficiency following transfection with lipofectamine 2000. The use of Fu-GENE HD, a non-liposomal based transfection reagent was demonstrated to provide 60% transfection efficiency with similar suppression of hCG- level as observed with lipofectamine 2000. The use of two transfection reagents to identify transfection efficiency and degree of suppression of gene expression hCG- was an appropriate step carried out by the authors. It has been demonstrated that selection of the most optimal carrier for gene transfer (which provides high transfection efficiency) is critical for expression of desired phenotype (Fortes et al., 2003). Polyethyleneimine (PEI) is a novel transfection reagent that has been demonstrated to bind to DNA with a high efficiency and provide an effective gene transfer (Ahn et al. 2008). It has been demonstrated that the cytotoxicity caused by transfection reagent can mask the effect of expression/inhibition of gene following transfection. Lipofectamine 2000 has been demonstrated to not significantly affect the viability of cells (Dalby et al., 2004). However, the inclusion of an only lipofectamine 2000 treated (mock transfected) cells would have provided a robust control and shown a more conclusive effect of snRNA against hCG on the viability of cells and apoptosis in Hela cells.
The next logical step for the authors would be evaluate the effectiveness of U1sRNA (anti-hCG-) in mouse tumor models of cervical carcinoma. Ye, et al. 2006 have established a mouse model for cervical cancer. Finally an efficient delivery system for delivery of the U1sRNA (anti-hCG-) has to be identified or developed for effective delivery and suppression of hCG- in vivo (mouse tumor) model of cervical cancer. Nanoparticle based technique has been utilized for encapsulation of siRNA or snRNA by using PEI or PLGA (Das et al., 2014). It might also be useful to design siRNA or shRNA sequences for hCG- and evaluate their efficacy in suppression of hCG- expression and inhibitory effect on cancer cell viability (Burczynska et al, 2013).
References
Ahn HH, Lee JH, Kim KS, Lee JY, Kim MS, Khang G, Lee IW, Lee HB (2008). Polyethyleneimine-mediated gene delivery into human adipose derived stem cells. Biomaterials. 29, (15):2415-22.
Burczynska, B., Booth, M. J., Iles, R. K., Shah, A., Shiled, A., & Butler, S. A. (2013). Stable Knockdown of hCGβ mRNA Expression in Bladder Cancer Cells Results in Significant Growth Inhibition. Anticancer research, 33(9), 3611-3614.
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Jankowska A, Gunderson SI, Andrusiewicz M, Burczynska B, Szczerba A, Jarmolowski A, Nowak-Markwitz E Warchol (2008). Reduction of human chorionic gonadotropin beta subunit expression by modified U1 snRNA caused apoptosis in cervical cancer cells. Molecular Cancer. 7:26
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