Analyses of Rac1 siRNA knockdown for nasopharyngeal carcinoma/HK1 cell line. Malaysian Journal of Medicine and Health Sciences, 15 (SPS). pp. 33-37. ISSN 2636-9346 (2019)
Abstract
Introduction: Patients with Nasopharyngeal carcinoma (NPC) usually diagnosed at advanced cancer stage and recurrent case. Rac1 have become an emerging therapeutic target for metastasis cancer. This gene is critically involved in cell polarization and reactive oxygen species-mediated cell killing. This study aims to investigate the Rac1 activities in NPC/HK1 cell line using siRNA approach and evaluate the calcium deposition profile.
Methods: The NPC/ HK1cells were transfected with Rac1-siRNA (siRac1) at concentrations of 50nM, 100nM and 200nM for 24 hours and stained with alizarin red s for calcium mineralization profile. Levels of Rac1 gene expression were measured via qRT-PCR followed by the time dependent assessment for 24, 48 and 72 hours.
Results: Findings revealed that siRac1 concentrations of 200nM (p-value <0.02) and 100nM (p-value <0.016) had significant Rac1 suppression while 50nM (p-value <0.076) had the least suppression. On the other hand, from alizarin red S staining showed no significant changes for calcium mineralization activity on treated and control cells. However, siRac1 treated cells at 200nM showed presence of intracellular organelle swelling and loss of membrane integrity in 70% of the cells. This observation could possibly be linked to early sign of necrosis activity, hypoxia and disruption in intracellular calcium influx.
Conclusion: This study suggest that Rac1 gene suppression might be involved in disruption of calcium deposition and reactive oxygen species-mediated NPC/HK1 cell killing. Further insight on the Rac1 molecular mechanism are needed to understand its potential role as therapeutic target for NPC.
| Item Type: | Article |
|---|---|
| Keywords: | Nasopharyngeal carcinoma, Rac1, Calcium mineralization in cancer, siRNA therapeutics, Reactive oxygen species-mediated cell killing |
| Taxonomy: | By Subject > Medicine > Pathology |
| Local Content Hub: | Subjects > Medicine |
| Depositing User: | Adi Azri Mohamad (Sg. Buloh) |
| Date Deposited: | 15 Feb 2022 23:40 |
| Last Modified: | 16 Feb 2022 02:54 |
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