Abstract
Abstract Integrin-linked kinase (ILK) is overexpressed in ovarian cancer cells. In the present study, we sought to assess the effect of ILK silencing on ovarian cancer cell proliferation using shRNA. Our lab previously demonstrated that phosphorylation of ILK acts as a proliferative signal in a variety of ovarian cancer cell lines. Cytotoxicity of ILK knockdown using shRNA was analyzed in vitro using SKOV3 cell lines. Transfected cells were incubated in Incucyte®, which allowed for simultaneous assessment of viable cells at the end of the incubation period using CellTiter glo assay. Knockdown of ILK using shRNA significantly inhibited the proliferation of SKOV3 cells. SKOV3 cells were then infected with lentivirus expressing ILK shRNA to obtain stable knockdown of ILK, which was confirmed with Western blot. Both parental and ILK shRNA infected cells were injected subcutaneously in NOD SCID Gamma immunocompromised mice. Tumor growth was assessed by sequential tumor measurements and final weights. Xenograft data indicated significantly decreased tumor volume over multiple time points in the transfected cells (p <0.0001) (Figure 1). Average final volume for the ILK-shRNA group (n = 15) and control group (n = 13) was 660mm3 (SD = 366.7, range 271 - 1667 mm3) and 1165 mm3 (SD = 668.7, range 506 - 2664 mm3), respectively. Similarly, average tumor weight was less for the ILK-shRNA group (404.4mg [range 226.1 - 801mg]) relative to control (584.3mg [range 293.2 - 1033.4mg]). Our findings demonstrate that knockdown of ILK in ovarian cancer cell lines results in decreased cell proliferation, as well as decreased tumor growth in xenograft models. This supports the role for further investigation into ILK as a potential therapeutic target in ovarian cancer. Days From Tumor Implantation1214161921232628313438VectorAverage Tumor Volume (mm^3)142.4151.6203274.7379.3482.1671.4741.2845.11083.81164.6Standard Error26.983.430.835.759.682112.3110.7133.9172.2185.5ILK-shRNAAverage Tumor Volume (mm^3)45.9107.282.4113150.9177263.4345.2397.3568.8659.9Standard Error6.39.99.915.225.822.841.558.455.19994.7 Citation Format: Tiffany Redfern, Benjamin Wilson, Michael Ulm, Adam ElNaggar, Suriyan Ponnusamy, Yinan Wang, Sarah Asemota, Ramesh Narayanan. Integrin-linked kinase gene knockdown results in decreased growth of ovarian cancer xenograft models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5044.
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