Increases in Doxorubicin Sensitivity and Radioiodide Uptake by Transfecting shMDR and Sodium/Iodide Symporter Gene in Cancer Cells Expressing Multidrug Resistance.
- Author:
Sohn Joo AHN
1
;
Yong Jin LEE
;
You La LEE
;
Chang Ik CHOI
;
Sang Woo LEE
;
Jeongsoo YOO
;
Byeong Cheol AHN
;
In Kyu LEE
;
Jaetae LEE
Author Information
1. Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu, Korea. jaetae@knu.ac.kr
- Publication Type:Original Article
- Keywords:
short hairpin RNA (shRNA);
Multidrug resistance;
Sodium/Iodide Symporter;
Tc-99m sestamibi;
Doxorubicin
- MeSH:
Adenoviridae;
Blotting, Western;
Colonic Neoplasms;
Doxorubicin*;
Drug Resistance, Multiple*;
Gene Expression;
HEK293 Cells;
Humans;
Ion Transport*;
Kidney;
Liposomes;
RNA, Messenger;
RNA, Small Interfering;
Transfection
- From:Nuclear Medicine and Molecular Imaging
2007;41(3):209-217
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Multidrug resistance (MDR) of the cancer cells related to mdr1 gene expression can be effectively treated by selective short hairpin RNA for mdr1 gene (shMDR). Sodium/iodide symporter (NIS) gene is well known to have both reporter and therapeutic gene characteristics. We have co-transfected both shMDR and NIS gene into colon cancer cells (HCT15 cell) expressing MDR and Tc-99m sestamibi and I-125 uptake were measured. In addition, cytotoxic effects of doxorubicin and I-131 therapy were also assessed after transfection. MATERIAL AND METHODS: At first, shMDR was transfected with liposome reagent into human embryonic kidney cells (HEK293) and HCT cells. shMDR transfection was confirmed by RT-PCR and western blot analysis. Adenovirus expressing NIS (Ad-NIS) gene and shMDR (Ad-shMDR) were co-transfected with Ad-NIS into HCT15 cells. Forty-eight hours after infection, inhibition of P-gycoprotein (Pgp) function by shMDR was analyzed by a change of Tc-99m sestamibi uptake and doxorubicin cytotoxicity, and functional activity of induced NIS gene expression was assessed with I-125 uptake assay. RESULTS: In HEK293 cells transfected with shMDR, mdr1 mRNA and Pgp protein expressions were down regulated. HCT15 cells infected with 20 MOI of Ad-NIS was higher NIS protein expression than control cells. After transfection of 300 MOI of Ad-shMDR either with or without 10 MOI of Ad-NIS, uptake of Tc-99m sestamibi increased up to 1.5-fold than control cells. HCT15 cells infected with 10 MOI of Ad-NIS showed approximately 25-fold higher I-125 uptake than control cells. Cotransfection of Ad-shMDR and Ad-NIS resulted in enhanced cytotoxic by doxorubicin in HCT15 cells. I-131 treatment on HCT15 cells infected with 20 MOI of Ad-NIS revealed increased cytotoxic effect. CONCLUSION: Suppression of mdr1 gene expression, retention of Tc-99m sestamibi, enhanced doxorubicin cytotoxicity and increases in I-125 uptake were achieved in MDR expressing cancer cell by co-transfection of shMDR and NIS gene. Dual therapy with doxorubicin and radioiodine after cotransfection shMDR and NIS gene can be used to overcome MDR.
