Ulm Michael A, Redfern Tiffany M, Wilson Ben R, Ponnusamy Suriyan, Asemota Sarah, Blackburn Patrick W, Wang Yinan, ElNaggar Adam C, Narayanan Ramesh
Division of Gynecologic Oncology, West Cancer Center and Research Institute, Memphis, TN 38138, USA.
Department of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
J Pers Med. 2020 Nov 26;10(4):246. doi: 10.3390/jpm10040246.
The objective of this study is to identify and validate novel therapeutic target(s) in ovarian cancer.
Development of targeted therapeutics in ovarian cancer has been limited by molecular heterogeneity. Although gene expression datasets are available, most of them lack appropriate pair-matched controls to define the alterations that result in the transformation of normal ovarian cells to cancerous cells.
We used microarray to compare the gene expression of treatment-naïve ovarian cancer tissue samples to pair-matched normal adjacent ovarian tissue from 24 patients. Ingenuity Pathway Analysis (IPA) was used to identify target pathways for further analysis. Integrin-linked kinase (ILK) expression in SKOV3 and OV90 cells was determined using Western blot. ILK was knocked down using CRISPR/Cas9 constructs. Subcutaneous xenograft study to determine the effect of ILK knockdown on tumor growth was performed in NOD SCID gamma mice.
Significant upregulation of the ILK pathway was identified in 22 of the 24 cancer specimens, identifying it as a potential player that could contribute to the transformation of normal ovarian cells to cancerous cells. Knockdown of ILK in SKOV3 cells resulted in decreased cell proliferation and tumor growth, and inhibition of downstream kinase, AKT (protein kinase B). These results were further validated using an ILK-1 chemical inhibitor, compound 22.
Our initial findings validate ILK as a potential therapeutic target for molecular inhibition in ovarian cancer, which warrants further investigation.
本研究的目的是识别和验证卵巢癌新的治疗靶点。
卵巢癌靶向治疗的发展受到分子异质性的限制。尽管有基因表达数据集，但其中大多数缺乏合适的配对对照来确定导致正常卵巢细胞转化为癌细胞的改变。
我们使用微阵列比较了24例未经治疗的卵巢癌组织样本与配对的正常相邻卵巢组织的基因表达。利用 Ingenuity 通路分析（IPA）来识别进一步分析的目标通路。使用蛋白质印迹法测定SKOV3和OV90细胞中整合素连接激酶（ILK）的表达。使用CRISPR/Cas9构建体敲低ILK。在NOD SCID gamma小鼠中进行皮下异种移植研究，以确定敲低ILK对肿瘤生长的影响。
在24个癌症标本中的22个中发现ILK通路显著上调，确定其为可能促成正常卵巢细胞转化为癌细胞的潜在因素。敲低SKOV3细胞中的ILK导致细胞增殖和肿瘤生长减少，并抑制下游激酶AKT（蛋白激酶B）。使用ILK-1化学抑制剂化合物22进一步验证了这些结果。
我们的初步研究结果验证了ILK作为卵巢癌分子抑制的潜在治疗靶点，值得进一步研究。
