Impact of Docetaxel conjugated Nanodiamonds on Gene expression of Prostate Cancer (PC3) Cells

J.D. Cedeño Ortiz, S. Joshi, S.R. Singh, K. Vig
Alabama State University,
United States

Keywords: docetaxel, PC3 cells, nanodiamonds, gene expression


Prostate cancer (PC) is the second leading cause of adult male death in the United States. This type of cancer is a disease where the prostate gland cells start to grow without control, changing in size and shape and provoking the development of malignant tumors that could lead to death. Docetaxel (Dox) drug is used to treat PC but it becomes toxic or builds resistance to cancer cells in patients. Possible treatments for PC suggested the use of loaded nanoparticles as therapeutics. In this study, nanodiamonds (ND’s) were used as carriers to deliver drug to PC cells. ND’s were conjugated with Docetaxel alone and along with RGD Peptide using EDC/NHS chemistry. To determine the location of ND’s in PC3 cells we prepared ND’s with NHS Fluorescein and tracked inside the cells by florescence microscopy. After 24 hours of incubation ND’s were located in the cell nucleus. A drug release assay was performed to estimate drug release profile from ND’s. It was found that 92.3% of drug was released in 24 hrs. Microculture Tetrazolium Assay (MTT) was done to determine the toxicity of ND’s and conjugated ND’s. After 48hrs of incubation, viability for HEp-2 and PC3 cells was 81% and 75% respectively at 50µg/mL of ND’s whereas it was 46% and 9% with ND + Dox and ND + Dox + RGD in PC3 stating that it efficient in causing cancer cell death. We did gene expression analysis on PC3 cells with ND’s and conjugated ND’s after 24 hours incubation. RNA was extracted using RNeasy Plus Mini Kit (Qiagen) and cDNA was synthesized, using the RT2 First Strand Kit (Qiagen). PCR Array was performed using a qPCR. Conjugated ND + Dox + RGD showed upregulation in many genes of PC cells. This study helped us know about how effective these nanoparticles are to deliver a drug into cancer cells and how efficient is the drug we tested to treat prostate cancer cells.