Zhawar Vikramjit K, Kandpal Raj P, Athwal Raghbir S
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA, U.S.A.
Department of Basic Medical Sciences, Western University of Health Sciences, Pomona, CA, U.S.A.
Cancer Genomics Proteomics. 2019 Jan-Feb;16(1):59-64. doi: 10.21873/cgp.20111.
The aberrant regulation of growth and proliferation is a key feature of carcinoma cells. In order to use molecular strategies to correct these defects toward therapeutic purposes, it is important to characterize the entire spectrum of causative molecules.
By using gene transfer technique, SKOV3 ovarian carcinoma cells were transduced with an expression construct of glutamate receptor 6 (glutamate ionotropic receptor kainate type subunit 2, GRIK2) in retroviral vector PQCXIP. The senescence of transduced cells was subsequently characterized.
Our results demonstrated that retroviral transduction occurs with high frequency and transduced cells continue to proliferate, albeit at a significantly reduced rate, up to 39 days. Some transduced colonies stopped proliferating after 12 days, and none of the clones proliferated beyond 37 days. The doubling time for these transduced cells increased progressively until they reached a complete cell-cycle arrest. The proliferating cells were distinguished by bromodeoxyuridine incorporation and 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay. The growth and cell cycle arrest in transduced cells accompanied activation of senescence-associated β-galactosidase. Furthermore, we have demonstrated a decrease in the levels of active protein kinase B and increase in the abundance of inactive cyclin-dependent kinase 1.
These results indicate involvement of GRIK2 in senescence and suggests GRIK2 as a potential target for therapeutic intervention of cancer cells.
生长和增殖的异常调节是癌细胞的一个关键特征。为了利用分子策略纠正这些缺陷以达到治疗目的，表征整个致病分子谱很重要。
通过基因转移技术，用逆转录病毒载体PQCXIP中的谷氨酸受体6（谷氨酸离子型受体红藻氨酸型亚基2，GRIK2）表达构建体转导SKOV3卵巢癌细胞。随后对转导细胞的衰老进行表征。
我们的结果表明，逆转录病毒转导高频发生，转导细胞持续增殖，尽管速率显著降低，长达39天。一些转导的集落在12天后停止增殖，没有克隆在37天后仍增殖。这些转导细胞的倍增时间逐渐增加，直到它们完全停滞于细胞周期。通过溴脱氧尿苷掺入和3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐测定法区分增殖细胞。转导细胞中的生长和细胞周期停滞伴随着衰老相关β-半乳糖苷酶的激活。此外，我们证明了活性蛋白激酶B水平降低，非活性细胞周期蛋白依赖性激酶1丰度增加。
这些结果表明GRIK2参与衰老，并提示GRIK2作为癌细胞治疗干预的潜在靶点。
