丁酸钠联合X射线照射对肺癌A549细胞增殖的抑制作用及其对细胞周期的影响

doi:10.13481/j.1671-587x.20210121

Abstract

Abstract: Objective

To study the inhibitory effect and radiosensitization effects of sodium butyrate （NaBt）， a histone deacetylase inhibitor （HDACi）， on the proliferation of human non-small cell lung cancer A549 cells alone or in combination with X-ray irradiation， and to analyze its combined effect with irradiation on the cell cycle of A549 cells.

Methods

The A549 cells in logarithmic growth phase were divided into control group， different concentrations （5， 10， 15， 20，and 25 mmol·L^－1） of NaBt groups， 4 Gy X-ray irradiation group and different concentrations （5， 10， 15， 20，and 25 mmol·L^－1） NaBt combined with 4 Gy X-ray irradiation groups. CCK-8 method was used to detect the cell proliferation rates in various groups at 24， 48 and 72 h after treatment. The changes of cell cycle were analyzed by flow cytometry.The A549 cells in logarithmic growth phase were divided into control group， 10 mmol·L^－1 NaBt group，20 mmol·L^－1NaBt group， 4 Gy X-ray irradiation group， 10 mmol·L^－1NaBt combined with 4 Gy X-ray irradiation group， and 20 mmol ·L^－1NaBt combined with 4 Gy X-ray irradiation group. At 24 h after treatment， the percentages of A549 cells in each phase of cell cycle in various groups were analyzed.

Results

After 48 h of treatment， the cell proliferation rates in 5，10， 15， 20， and 25 mmol·L^－1 NaBt groups were significantly lower than that in control group（P<0.05 or P<0.01）；after 72 h of treatment， the cell proliferation rates in 5，10，20，and 25 mmol·L^－1 NaBt groups were significantly lower than that in control group（P<0.01） in a concentration- and time-dependent manner；after treated for 48 h， the cell proliferation rates in 15， 20，and 25 mmol·L^－1 NaBt combined with 4 Gy X-ray irradiation groups were significantly lower than those in control group，4 Gy X-ray irradiation group and corresponding concentrations of NaBt groups （P<0.05 or P<0.01）；after treated for 72 h，compared with control group and 4 Gy X-ray irradiation group，the cell proliferation rates in different concentrations of NaBt combined with 4 Gy X-ray irradiation groups were significantly decreased（P<0.01）；the cell proliferation rates in 20 and 25 mmol·L^－1 NaBt combined with 4 Gy X-ray irradiation groups were significantly lower than those in corresponding concentrations of NaBt groups（P<0.01）.The cell cycle analysis results showed that after 24 h of treatment， compared with control group， the percentages of A549 cells in G₀ / G₁ phase and G₂+ M phase in 10 and 20 mmol·L^－1 NaBt groups were significantly increased （P<0.05 or P<0.01）， and the percentage of A459 cells in S phase was significantly decreased （P<0.01）； the percentages of cells in G₂+M phase in 4 Gy X-ray irradiation group and 10 mmol·L^－1NaBt combined with 4 Gy X-ray irradiation group were significantly increased （P<0.05 or P<0.01），and the percentages of cells in G₀/G₁ phase and S phase were significantly decreased （P<0.01）.

Conclusion

NaBt has the inhibitory effect on the proliferation of A549 cells and radiosensitization effect in a concentration- and time-dependent manner， and the combined inhibitory effect of NaBt and X-ray is better than both of them used alone； its mechanism may be related to G₂+ M phase arrest of the A549 cells induced by combination of NaBt and X-ray.

Key words: histone deacetylase inhibitor, sodium butyrate, non- small cell lung cancer, epigenetics, radiosensitivity

CLC Number:

Q691

Guanhu LI,Qingxu LANG,Chunyan LIU,Qin LIU,Mengrou GENG,Xiaoqian LI,Zhenqi WANG. Inhibitory effect of sodium butyrate combined with X- irradiation on proliferation of lung cancer A549 cells and its influence in cell cycle[J].Journal of Jilin University(Medicine Edition), 2021, 47(1): 152-157.

Figures/Tables 2

Tab.1

Tab.2

References 24

1	马书梅，刘瑞娟，张影影. 肺癌的流行病学［J］. 世界最新医学信息文摘， 2019， 19（26）： 77-78， 80.
2	YASMIN R， SIRAJ S， HASSAN A， et al. Epigenetic regulation of inflammatory cytokines and associated genes in human malignancies ［J］. Mediators Inflamm， 2015， 2015：201703.
3	SETO E， YOSHIDA M. Erasers of histone acetylation： the histone deacetylase enzymes ［J］. Cold Spring Harb Perspect Biol， 2014， 6（4）： a018713.
4	OKUDELA K， MITSUI H， SUZUKI T， et al. Expression of HDAC9 in lung cancer：potential role in lung carcinogenesis ［J］. Int J Clin Exp Pathol， 2014， 7（1）： 213-220.
5	FALKENBERG K J， JOHNSTONE R W. Histone deacetylases and their inhibitors in cancer， neurological diseases and immune disorders ［J］. Nat Rev Drug Discov， 2014， 13（9）： 673-691.
6	SURAWEERA A， O’BYRNE K J， RICHARD D J， et al. Combination therapy with histone deacetylase inhibitors （HDACi） for the treatment of cancer： achieving the full therapeutic potential of HDACi ［J］. Front Oncol， 2018， 8：92.
7	MCMILLAN M T， OJERHOLM E， VERMA V， et al. Radiation treatment time and overall survival in locally advanced non-small cell lung cancer ［J］. Int J Radiat Oncol Biol Phys， 2017， 98（5）： 1142-1152.
8	HESSMANN E， JOHNSEN S A， SIVEKE J T， et al. Epigenetic treatment of pancreatic cancer： Is there a therapeutic perspective on the horizon？［J］. Gut， 2017， 66（1）：168-179.
9	BAO L M，DIAO H，DONG N， et al. Histone deacetylase inhibitor induces cell apoptosis and cycle arrest in lung cancer cells via mitochondrial injury and p53 up-acetylation ［J］. Cell Biol Toxicol， 2016， 32（6）： 469-482.
10	FISCHER C， LEITHNER K， WOHLKOENIG C， et al. Panobinostat reduces hypoxia-induced cisplatin resistance of non-small cell lung carcinoma cells via HIF-1α destabilization ［J］. Mol Cancer， 2015， 14： 4.
11	TO K K， TONG W S， FU L W. Reversal of platinum drug resistance by the histone deacetylase inhibitor belinostat ［J］. Lung Cancer， 2017， 103： 58-65.
12	MINAMIYA Y， ONO T， SAITO H， et al. Expression of histone deacetylase 1 correlates with a poor prognosis in patients with adenocarcinoma of the lung［J］. Lung Cancer， 2011， 74（2）： 300-304.
13	OSADA H， TATEMATSU Y， SAITO H， et al. Reduced expression of class Ⅱ histone deacetylase genes is associated with poor prognosis in lung cancer patients ［J］. Int J Cancer， 2004， 112（1）： 26-32.
14	ZHANG Q C， JIANG S J， ZHANG S， et al. Histone deacetylase inhibitor trichostatin A enhances anti-tumor effects of docetaxel or erlotinib in A549 cell line ［J］. Asian Pac J Cancer Prev， 2012， 13（7）： 3471-3476.
15	GROSELJ B， SHARMA N L， HAMDY F C， et al. Histone deacetylase inhibitors as radiosensitisers： effects on DNA damage signalling and repair ［J］. Br J Cancer， 2013， 108（4）： 748-754.
16	ADIMOOLAM S， SIRISAWAD M， CHEN J， et al. HDAC inhibitor PCI-24781 decreases RAD51 expression and inhibits homologous recombination ［J］. Procl Natl Acad Sci USA， 2007， 104 （49）： 19482-19487.
17	MCMILLAN M T， OJERHOLM E， VERMA V， et al. Radiation treatment time and overall survival in locally advanced non-small cell lung cancer ［J］. Int J Radiat Oncol Biol Phys， 2017， 98（5）： 1142-1152.
18	JIN X F， WU N N， DAI J J， et al. TXNIP mediates the differential responses of A549 cells to sodium butyrate and sodium 4-phenylbutyrate treatment ［J］. Cancer Med，2017， 6（2）： 424-438.
19	LI L， SUN Y， LIU J， et al. Histone deacetylase inhibitor sodium butyrate suppresses DNA double strand break repair induced by etoposide more effectively in MCF-7 cells than in HEK293 cells ［J］. BMC Biochem， 2015， 16（1）： 2.
20	BOURKE E， BROWN J A，TAKEDA S， et al. DNA damage induces Chk1-dependent threonine-160 phosphorylation and activation of Cdk2 ［J］. Oncogene， 2010， 29（4）： 616-624.
21	敖经盛, 鲍志荣, 全红. 纳米复合材料GO@AgPt对非小细胞肺癌的放射增敏作用[J].中国医学物理学杂志,2020,37(4): 514-519.
22	TAILOR D，HAHM E R，KALE R K， et al. Sodium butyrate induces DRP1-mediated mitochondrial fusion and apoptosis in human colorectal cancer cells ［J］. Mitochondrian， 2014， 16： 55-64.
23	MAHA S A， KAREM H A， SAIED A J， et al. The effect of combined treatment with sodium phenylbutyrate and cisplatin， erlotinib， or gefitinib on resistant NSCLC cells ［J］. Clin Pharmacol， 2018，10： 135-140.
24	RIVERA S，LETEUR C，MEGNIN F， et al. Time dependent modulation of tumor radiosensitivity by a pan HDAC inhibitor： abexinostat ［J］. Oncotarget， 2017， 8（34）： 56210-56227.

Group	Proliferation rate
Group	（t/h） 24	48	72
Control	1.00±0.00	1.00±0.00	1.00±0.00
NaBt (mmol·L^－1)
5	0.95±0.33	0.95±0.01^**	0.61±0.01^**
10	0.98±0.32	0.98±0.01	0.51±0.04^**
15	0.98±0.28	0.96±0.01^*	0.58±0.02^**
20	1.00±0.03	0.95±0.01^**	0.62±0.01^**
25	0.96±0.04	0.91±0.02^**	0.53±0.03^**
4 Gy X-ray irradiation	1.02±0.01	0.98±0.01	0.96±0.05
NaBt+4 Gy X-ray irradiation
5 mmol·L^－1+4 Gy	1.06±0.02^*	0.99±0.03	0.61±0.03^**##
10 mmol·L^－1+4 Gy	1.15±0.06^*	0.99±0.04	0.53±0.04^**##
15 mmol·L^－1+4 Gy	1.08±0.01^**	0.91±0.02^**△##	0.55±0.03^**##
20 mmol·L^－1+4 Gy	1.06±0.02^**	0.91±0.02^**△#	0.49±0.01^**△△##
25 mmol·L^－1+4 Gy	1.02±0.01	0.89±0.01^**△##	0.39±0.02^**△△##

Group	Percentage of A549 cells
Group	G₀/G₁	G₂ + M	S
Control	83.057 ± 0.071	2.060 ± 0.839	14.883±0.783
NaBt (mmol·L^－1) 10	93.203±0.482^**	3.923±0.245	2.873±0.062^**
20	88.890±0.538^**	7.287±0.345^*	3.823±0.049^**
4 Gy X-ray irradiation	69.103±0.501^**	27.490±0.540^**	3.403±0.057^**
10 mmol·L^－1NaBt +4 Gy X-ray irradiation	64.133±0.904^**	32.593±0.952^**	3.270±0.162^**
20 mmol·L^－1NaBt +4 Gy X-ray irradiation	—	—	—

[1]	XIN Rui, LI Dong, XU Huiying, WANG Chunyu, QU Danhua, SUN Haifeng. Influence of miRNA-21 in radiosensitivity of breast cancer cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 772-778.
[2]	YANG Wenyan, LIU Qiang, SUN Zhijuan, DU Liqing, XU Chang, WANG Yan, LIU Yang, WANG Qin. Effect of melatonin on radiosensitivity of non-small cell lung cancer H1299 cells [J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 532-536.
[3]	TU Peipei, LI Xiaodan, MA Baicheng, ZHANG Yaofang, DUAN Huikun, NI Zaizhong, WANG Haisong, JIANG Pingzhe, LI Miao, WU Ri, LI Minggang. Changes and significance of hepatic histone H3 epigenetic modification in type 2 diabetic mice [J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 756-762.
[4]	YANG Yan-ming,FANG Fang,LIU Nian,QU Zhi-jie，WANG Zhi-cheng,ZHANG Yan-qiu. Influence of pEgr1-hsTRAL plasmid on radiosensitivity and DR4 and 　DR5 expression levels in lung adenocarcinoma A549 cells [J]. Journal of Jilin University Medicine Edition, 2013, 39(2): 199-203.
[5]	LIANG Bing,LIU Xiao-dong,XU Hui-ying,XIN Rui,KONG De-juan,HE Meng-zi,JIA Li-li,. Effects of deoxycytidine kinase on sensitivity to radiotherapy and chemotherapy in HeLa cells  [J]. J4, 2012, 38(2): 197-201.
[6]	ZHAO Tong, WANG Xiu-Lan, LIU Yong, WANG Ruo-Yu. Radiosensitization of epithelial growth factor receptor monoclonal antibody on lung adenocarcinoma cell line SPC-A-1 [J]. J4, 2011, 37(3): 483-486.
[7]	SUN Bao-sheng,LIU Shi-xin,WANG Tie-jun,HUANG Guo-min,GONG Shou-liang,LIU Lin-lin. Effects of taurolidine on radiosensitivity of murine melanoma cells and its mechanism [J]. J4, 2008, 34(6): 923-926.
[8]	DI Qian, LI Yang, ZHAO Xue-jian, MIYAGI Taeko. Effects of sodium butyrate on cell growth inhibitionand apoptosis in PC-3M cell lines [J]. J4, 2004, 30(5): 710-712.

Inhibitory effect of sodium butyrate combined with X- irradiation on proliferation of lung cancer A549 cells and its influence in cell cycle

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