丙戊酸联合X射线照射对乳腺癌MDA-MB-231细胞增殖的抑制作用及其机制

doi:10.13481/j.1671-587X.20220310

Abstract

Abstract: Objective

To observe the inhibitory effect of valproic acid （VPA）， a histone deacetylase inhibitor （HDACi）， on the proliferation of the human triple negative breast cancer （TNBC）MDA-MB-231 cells alone or combined with X-ray irradiation， and to analyze the effect of combined irradiation on the apoptosis and autophage of the MDA-MB-231 cells.

Methods

The MDA-MB-231 cells at logarithmic growth phase were divided into control group， different concentrations（2.5，5.0，10.0，20.0， and 40.0 mmol·L^－1） of VPA groups 4 Gy， X-ray irradiation groups，and different concentrations （2.5，5.0，10.0，20.0， and 40.0 mmol·L^－1） of VPA combined with 4 Gy X-ray irradiation groups. The proliferation rates of the cells in various groups were detected by CCK-8 method after treated for 24， 48， and 72 h.The cells were divided into control group， 4 Gy X-ray irradiation group， 5 mmol·L^－1 VPA group， 5 mmol·L^－1 VPA combined with 4 Gy X-ray irradiation group， the apoptotic rates and autophage rates of the cells in various groups were analyzed by fiow cytometry after treated for 24 and 48 h.

Results

Compared with control group， the proliferation rates of the MDA-MB-231 cells in VPA group and VPA combined with 4 Gy X-ray irradiation groups were decreased significantly in a concentration and time-dependent manner （P<0.01）. Compared with 4 Gy X-ray irradiation group， the proliferation rates of the MDA-MB-231cells in VPA combined with 4 Gy X-ray irradiation groups were significantly decreased （P<0.01）. Compared with 2.5 mmol·L^－1 VPA group， the proliferation rate of the MDA-MB-231 cells in 2.5 mmol·L^－1 VPA combined with 4 Gy X-ray irradiation group after treated for 24 h was decreased（P<0.05）. After treated for 72 h， the proliferation rates of the MDA-MB-231 cells in 2.5 and 10.0 mmol·L^－1 VPA combined with 4 Gy X-ray irradiation groups were lower than those in 2.5 and 10.0 mmol·L^－1 VPA groups（P<0.05），respectively. After treated for 24 and 48 h， the apoptotic rates and autophagy rates of the MDA-MB-231 cells in 4 Gy X-ray group， 5 mmol·L^－1 VPA group and 5 mmol·L^－1 VPA combined with 4 Gy X-ray group were significantly higher than those in control group （P<0.05 or P<0.01）. Compared with 4 Gy X-ray irradiation group， the apoptotic rate and autophagy rate of the MDA-MB-231 cells in 5 mmol·L^－1 VPA combined with 4 Gy X-ray irradiation group were significantly increased （P<0.01）. After treated for 48 h，compared with 5 mmol·L^－1 VPA group，the autophagy rate of the MDA-MB-231 cells in 5 mmol·L^－1 VPA combined with 4 Gy X-ray irradiation group was increased （P<0.05）.

Conclusion

VPA can inhibit the proliferation of MDA-MB-231 cells in a concentration and time-dependent manner， and the combined inhibitory effect of VPA and X-ray is better， and the mechanism may be involved in the increasing of apoptosis and autophage of the MDA-MB-231 cells induced by the combination of VPA and X-ray.

Key words: Histone deacetylase inhibitor, Valproic acid, Cell proliferation inhibition, cell apoptosis, Autophage

CLC Number:

R736.3

Guanhu LI,Qingxu LANG,Chunyan LIU,Qin LIU,Mengrou GENG,Xiaoqian LI,Zhenqi WANG. Inhibitory effect of valproic acid combined with X-ray irradiation on proliferation of breast cancer MDA-MB-231 cells and its mechanism[J].Journal of Jilin University(Medicine Edition), 2022, 48(3): 622-629.

Figures/Tables 4

Tab. 1

Fig. 1

Tab. 2

Fig. 2

References 29

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Group	Proliferation rate
Group	（t/h） 24	48	72
Control	1.00 ± 0.00	1.00 ± 0.00	1.00 ± 0.00
4 Gy X-ray irradiation	0.92 ± 0.04	1.00 ± 0.01	0.56 ± 0.02^**
VPA ( mmol·L^－1)
2.5	0.73±0.04^*△	0.66±0.04^*△	0.53 ± 0.03^*
5.0	0.47±0.03^*△	0.48±0.05^*△	0.36 ± 0.05^*△
10.0	0.28±0.01^*△	0.33±0.03^*△	0.23 ± 0.01^*△
20.0	0.24±0.01^*△	0.24±0.00^*△	0.14 ± 0.00^*△
40.0	0.25±0.01^*△	0.25±0.00^*△	0.14 ± 0.00^*△
VPA (mmol·L^－1)+4 Gy X-ray irradiation
2.5 mmol·L^－1 VPA+4 Gy X-ray irradiation	0.632 ± 0.042^*△＃	0.68 ± 0.02^*△	0.41 ± 0.03^*△＃
5.0 mmol·L^－1 VPA+4 Gy X-ray irradiation	0.576 ± 0.024^*△	0.54 ± 0.04^*△	0.35 ± 0.01^*△
10.0 mmol·L^－1 VPA+4 Gy X-ray irradiation	0.339 ± 0.020^*△	0.40 ± 0.03^*△	0.22 ± 0.00^*△○
20.0 mmol·L^－1 VPA+4 Gy X-ray irradiation	0.271 ± 0.009^*△	0.25 ± 0.00^*△	0.14 ± 0.00^*△
40.0 mmol·L^－1 VPA+4 Gy X-ray irradiation	0.260 ± 0.003^*△	0.26 ± 0.01^*△	0.14 ± 0.00^*△

Group	Apoptotic rate		Autophagy rate
Group	24 h	48 h	24 h	48 h
Control	6.27. ± 0.53	6.46 ± 1.26	8.17 ± 0.74	7.73 ± 0.62
4 Gy X-ray irradiation	10.73 ± 0.81^**	22.50 ± 2.27^**	8.56 ± 0.49	32.60 ± 2.43^**
5.0 mmol·L^－1 VPA	41.24 ± 6.22^*△	57.91 ± 12.03^*△	42.17 ± 3.23^**△△	30.27 ± 1.70^**
5.0 mmol·L^－1 VPA +4 Gy X-ray irradiation	42.39 ± 4.15^**△△	58.24 ± 3.88^**△△	38.60 ± 5.49^*△	40.63 ± 0.65^**△＃

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Inhibitory effect of valproic acid combined with X-ray irradiation on proliferation of breast cancer MDA-MB-231 cells and its mechanism

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