Construction of Recombinant Vector on IL-24 and Its Effect in Combination with Chemotherapy Drugs for TUBB3 and ERCC-1 Expression in SKOV3 Cells†

doi:10.7503/cjcu20140975

Abstract

Abstract:

Gene fragments of IL-24 were cloned into pcDNA3.0-OSP-1-IL-24 and pcDNA3.0-IL-24 vector using gene recombination technology, the expression of two vectors was measured by RT-PCR methods. The two chemotherapy drugs of paclitaxel and cisplatin were applied into normal SKOV3 cells, stable transfected SKOV3 cells of pcDNA3.0, PcDNA3.0-IL-24, pcDNA3.0-OSP-1 and pcDNA3.0-OSP-1-IL-24, respectively. The cell killing effect and proliferation ability of IL-24 in combination with chemotherapy drugs were detected by 3-(4,5)-dimethylthiahiahiazo-(z-yl)-2,5-di-phenytetrazolium bromide(MTT) in SKOV3 cells. Gene expression of TUBB3 and ERCC-1 were detected by real-time fluorescent quantitative polymerase chain reaction(PCR) technology in cell of each group. RT-PCR results showed that the pcDNA3.0-IL-24 and pcDNA3.0-OSP-1-IL-24 vector were cloned successfully by gene recombination technology. IL-24 gene can increase the sensitivity of SKOV3 cells to chemotherapeutic drug of cisplatin and paclitaxel. Its combination with chemotherapy drugs for IC₅₀ of tumor drugs decreased 10 times and 6 times, respectively. Gene expression of TUBB3 gene and ERCC-1 decreased 4 times and 10 times, respectively. These results suggested that IL-24 in combination with chemotherapy drugs could reduce gene expression of TUBB3 and ERCC-1. This experiment provided an important theory basis for the clinical treatment of ovarian cancer.

Key words: Interleukin-24, Cisplatin, Paclitaxel, SKOV3 cell

CLC Number:

O629
R737.31

TrendMD:

ZHU He, DU Zhenwu, FAN Limei, GAO Haicheng, CUI Manhua. Construction of Recombinant Vector on IL-24 and Its Effect in Combination with Chemotherapy Drugs for TUBB3 and ERCC-1 Expression in SKOV3 Cells^†[J]. Chem. J. Chinese Universities, 2015, 36(5): 927.

Figures/Tables 5

References 20

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Gene	Primer sequence	Base pair
ERCC-1	F: 5'-ACGCCGAATATGCCATCTCAC-3'	292 bp
	R: 5'-AGCCGCCCATGGATGTAGTCT-3'
TUBB3	F: 5'-TTCCAGCTGACCCACTCTCT-3'	226 bp
	R: 5'-ACAGGGCCTCGTTATCAATG-3'
GAPDH	F: 5'-CCACATCGCTCAGACACCAT-3'	196 bp
	R: 5'-ACGGTGCCATGGAATTTGCC-3'

Gene	Primer sequence	Base pair
ERCC-1	F: 5'-ACGCCGAATATGCCATCTCAC-3'	292 bp
	R: 5'-AGCCGCCCATGGATGTAGTCT-3'
TUBB3	F: 5'-TTCCAGCTGACCCACTCTCT-3'	226 bp
	R: 5'-ACAGGGCCTCGTTATCAATG-3'
GAPDH	F: 5'-CCACATCGCTCAGACACCAT-3'	196 bp
	R: 5'-ACGGTGCCATGGAATTTGCC-3'

Drug	IC₅₀/(μg·mL^-1)
Drug	SKOV3 cell	pcDNA3.0- SKOV3 cell	pcDNA3.0-OSP-1- SKOV3 cell	pcDNA3.0-IL-24- SKOV3 cell	pcDNA3.0-OSP-1- IL-24- SKOV3 cell
DDP	5.760±0.560	5.012±0.263	5.165±0.564	0.505±0.032^*	0.567±0.041^*
PTX	12.760±1.870	11.452±2.852	10.989±1.568	2.120±0.690^*	3.142±0.041^*

Drug	IC₅₀/(μg·mL^-1)
Drug	SKOV3 cell	pcDNA3.0- SKOV3 cell	pcDNA3.0-OSP-1- SKOV3 cell	pcDNA3.0-IL-24- SKOV3 cell	pcDNA3.0-OSP-1- IL-24- SKOV3 cell
DDP	5.760±0.560	5.012±0.263	5.165±0.564	0.505±0.032^*	0.567±0.041^*
PTX	12.760±1.870	11.452±2.852	10.989±1.568	2.120±0.690^*	3.142±0.041^*

Gene	IC₅₀/(μg·mL^-1)
Gene	SKOV3 cell	pcDNA3.0- SKOV3 cell	pcDNA3.0-OSP-1- SKOV3 cell	pcDNA3.0-IL-24- SKOV3 cell	pcDNA3.0-OSP-1- IL-24-SKOV3 cell
ERCC1	1.240±0.230	2.012±0.293	1.165±0.574	0.112±0.023^*	0.141±0.027^*
TUBB3	1.540±0.130	1.452±0.872	1.989±0.569	0.426±0.023^*	0.445±0.074^*