Effect of miRNA-125b-1 on radiosensitivity of SKBR-3 breast cancer cells

doi:10.3877/cma.j.issn.1674-0807.2011.06.007

Abstract

Abstract:

Objective

To investigate the effect of miRNA-125b-1 on the radiosensitivity of SKBR-3 breast cancer cells, and its influence on HER-2 protein expression.

Methods

A recombinant miRNA-125b-1 was introduced by lipofectamine^TM 2000 mediated gene transfection into cultured SKBR-3 cells. The transfected cell clones were collected and analyzed by RT-PCR to determine the level of HER-2 mRNA. The protein level of HER-2 was detected by Western blot. The cell cycle phases were determined by flow cytometry, and the cell proliferation inhibitory rate was measured using MTT assay. The surviving fraction of the cells was examined by colony forming assay after the irradiation of different doses of X-ray. A single-hit, multi-target mathematic model was established to estimate the values of D₀, D_q and N. The radiosensitivity of cells was compared accordingly.One-way ANOVA was used for comparison of the results by RT-PCR and Western blot between groups,and ANOVA of repeated measurement data was carried out for comparison of cell cycle phases and cell proliferation between groups.

Results

In the experiment group,both the mRNA and protein expression levels of HER-2 gene in the transfected SKBR-3 cells were reduced significantly(all P＜0.05). Most of the transfected SKBR-3 cells were blocked in G₂M phase of cell cycle. The cell proliferation inhibitory rate was obviously higher than that in the negative control group (P＜0.05). The values of D₀, D_q and N were obtained through the cell survival curves. The values of transfected SKBR-3 cells (D₀=1.87,D_q=2.68,N=4.21) were significantly lower than those in the control group(D₀ =2.51,D_q=4.10,N=5.11;SER=1.34,P＜0.05), which demonstrated that the radiosensitivity of these cells was enhanced.

Conclusion

miRNA-125b-1 can markedly increase the radiosensitivity of SKBR-3 breast cancer cells,which inspires a new treatment for HER-2 positive breast cancer.

Key words: breast neoplasms, radiotherapy, miRNA-125b-1, microRNA, HER-2

Bin HU, Ai-guo WU, Xue-xiao LI, Shu-feng JI, Meng-chuan WANG, Kai WU, Guo-li SHAO. Effect of miRNA-125b-1 on radiosensitivity of SKBR-3 breast cancer cells[J]. Chinese Journal of Breast Disease(Electronic Edition), 2011, 05(06): 703-712.

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Figures/Tables 9

References 23

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组别	HER-2 mRNA表达量	F值	P值
脂质体对照组	0.82±0.00^a	447.78	0.00
阴性对照组	0.81±0.00^b
miRNA-125b-1组	0.66±0.01

组别	HER-2 mRNA表达量	F值	P值
脂质体对照组	0.82±0.00^a	447.78	0.00
阴性对照组	0.81±0.00^b
miRNA-125b-1组	0.66±0.01

组别	HER-2 mRNA表达量	F值	P值
脂质体对照组	0.79±0.05^a	10.07	0.01
阴性对照组	0.59±0.13^b
miRNA-125b-1组	0.36±0.15

组别	HER-2 mRNA表达量	F值	P值
脂质体对照组	0.79±0.05^a	10.07	0.01
阴性对照组	0.59±0.13^b
miRNA-125b-1组	0.36±0.15

组别	G₀-G₁期	G₂M期	S期
脂质体对照组	67.22±7.30	8.07±1.73	24.71±6.41
阴性对照组	64.56±6.88	10.42±1.47	25.02±5.66
miRNA-125b-1组	50.56±0.93	23.20±1.07	26.13±0.93

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