[an error occurred while processing this directive] | [an error occurred while processing this directive]
Effect of HMGB1 knockdown by RNA interference on cell proliferation and migration of esophageal squamous cell carcinoma after X-ray radiation
Yang Xingxiao1, Zhang Xueyuan2, Zou Naiyi2, Ma Ming3, Zhu Shuchai2
1Department of Infection,Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China; 2Department of Radiation Oncology, Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China; 3Department of Laboratory,Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China
AbstractObjective To evaluate the effect of X-ray radiation on cell proliferation, migration, survival ability and cell cycle of human esophageal squamous cell carcinoma after RNA interference-mediated down-regulation of HMGB1 gene expression. Methods The expression of HMGB1 at mRNA and protein levels in the human esophageal squamous cell carcinoma cell lines ECA109 and KYSE30 was determined using RT-PCR and Western blot assays. MTS and Transwell assays were employed to examine the proliferation and migration of ECA109 and KYSE30 cell lines. The cellular survival ability in vitro was assessed by clone formation assay. The cell cycle after X-ray radiation in different groups was detected by flow cytometry. Results The expression of HMGB1 at mRNA and protein levels in ECA109 and KYSE30 cells were markedly higher in a dose-dependent and time-dependent manner in the radiation group than that in the control group (all P<0.05). MTS results demonstrated that the proliferation of ECA109 and KYSE30 cells was obviously lower at each time point after radiation than that in the group without radiation (all P<0.01). The expression of HMGB1 at mRNA and protein levels was significantly inhibited in the HMGB1 siRNA group than those in the control and NC groups (both P<0.01). The data from the clone formation assay revealed that the radiosensitivity was significantly increased after down-regulation of HMGB1 expression (P<0.01). Transwell migration assay revealed that the number of migrating cells at the fourth hour after X-ray irradiation in the HMGB1 siRNA group was significantly lower than those in the control and negative groups (both P<0.01). In the HMGB1 siRNA group, the percentage of cells at G0/G1 phase was obviously higher, whereas the percentage of S phase was significantly lower than those in the control and NC groups, and the trend was even more significant after X-ray radiation (all P<0.01). Conclusion Inhibition of HMGB1 expression by siRNA can suppress the proliferation and migration of ECA109 and KYSE30 cells and enhance the radiosensitivity by increasing the cell cycle arrest at G0/G1 stage after X-ray irradiation in vitro.
Fund:National Natural Science Foundation of China (81872456;81903118);National Natural Science Foundation of Hebei Province (H2017206170);Hebei Institute of Medical Sciences (20160183,20170154,20180483)
Yang Xingxiao,Zhang Xueyuan,Zou Naiyi et al. Effect of HMGB1 knockdown by RNA interference on cell proliferation and migration of esophageal squamous cell carcinoma after X-ray radiation[J]. Chinese Journal of Radiation Oncology, 2019, 28(12): 933-938.
Yang Xingxiao,Zhang Xueyuan,Zou Naiyi et al. Effect of HMGB1 knockdown by RNA interference on cell proliferation and migration of esophageal squamous cell carcinoma after X-ray radiation[J]. Chinese Journal of Radiation Oncology, 2019, 28(12): 933-938.
[1]Siegel R, Ma J, Zou Z,et al. Cancer Statistics, 2014[J]. CA Cancer J Clin,2014,64(1):9-29. DOI:10.3322/caac.21208. [2]Siegel R,Naishadham D,Jemal A. Cancer statistics for Hispanics/Latinos,2012[J]. CA Cancer J Clin,2012,62(5):283-298. DOI:10.3322/caac.21153. [3]Meng Q,Zhao J,Liu H,et al. HMGB1 promotes cellular proliferation and invasion,suppresses cellular apoptosis in osteosarcoma[J]. Tumour Biol,2014,35(12):12265-12274. DOI:10.1007/s13277-014-2535-3. [4]Livak KJ,Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) method[J]. Methods,2001,25(4):402-408. DOI:10.1006/meth.2001.1262. [5]Chen J,Xi B,Zhao Y,et al. High-mobility group protein B1(HMGB1) is a novel biomarker for human ovarian cancer[J]. Gynecol Oncol,2012,126(1):109-117. DOI:10.1016/j.ygyno.2012.03.051. [6]Huang Z,Zhong Z,Zhang L,et al. Down-regulation of HMGB1 expression by shRNA constructs inhibits the bioactivity of urothelial carcinoma cell lines via the NF-κB pathway[J]. Sci Rep,2015,5:12807. DOI:10.1038/srep12807. [7]Chen RC,Yi PP,Zhou RR,et al. The role of HMGB1-RAGE axis in migration and invasion of hepatocellular carcinoma cell lines[J]. Mol Cell Biochem,2014,390(1/2):271-280. DOI:10.1007/s11010-014-1978-6. [8]Liu W,Zhang Z,Zhang Y,et al. HMGB1-mediated autophagy modulates sensitivity of colorectal cancer cells to oxaliplatin via MEK/ERK signaling pathway[J]. Cancer Biol Ther,2015,16(4):511-517. DOI:10.1080/15384047.2015.1017691. [9]Chen J,Liu X,Zhang J,et al. Targeting HMGB1 inhibits ovarian cancer growth and metastasis by lentivirus-mediated RNA interference[J]. J Cell Physiol,2012,227(11):3629-3638. DOI:10.1002/jcp.24069. [10]Bald T,Quast T,Landsberg J,et al. Ultraviolet-radiation-induced inflammation promotes angiotropism and metastasis in melanoma[J]. Nature,2014,507(7490):109-113. DOI:10.1038/nature13111. [11]张晓娟,王慧娟,张米娜,等. HMGB1/TLR4通路参与Fibulin-5对肺癌细胞增殖和转移的抑制作用[J]. 中国癌症杂志,2017,27(10):761-769. DOI:10.19401/j.cnki.1007-3639.2017.10.001. Zhang XJ,Wang HJ,Zhang MN,et al. HMGB1/TLR4 pathway s/HSbin-5 inhibition of lung cancer cell proliferation and metastasis[J]. Chin Oncol,2017,27(10):761-769. DOI:10.19401/j.cnki.1007-3639.2017.10.001. [12]肖威,华胜妮,肖芦山,等. 高迁移率组蛋白1对人鼻咽癌细胞株C666-1体外迁移及侵袭能力的影响[J]. 肿瘤防治研究,2015,42(10):955-959. DOI:10.3971/j.issn.1000-8578.2015.10.001. Xiao W,Hua SN,Xiao LS,et al. High migration rate histone serotonin 1 effect on in vitro migration and invasive ability of human nasopharyngeal cancer cell strains C666-1[J]. Chin J Cancer Prev Treat,2015,42(10):955-959. DOI:10.3971/j.issn.1000-8578.2015.10.001. [13]Meng Q,Zhao J,Liu H,et al. HMGB1 promotes cellular proliferation and invasion,suppresses cellular apoptosis in osteosarcoma[J]. Tumour Biol,2014,35(12):12265-12274. DOI:10.1007/s13277-014-2535-3. [14]印胡滨,何卫阳,苟欣,等.siRNA干扰HMGB1表达对膀胱癌T24株细胞恶性生物学行为的影响及其可能的机制[J]. 中国肿瘤生物治疗杂志,2017,24(3):264-270. DOI:10.3872/j.issn.1007-385X.2017.03.009. In HB,He WY,Gou X,et al.siRNA interference HMGB1 expresses its effect on the malignant biological behavior of bladder cancer T24 cells and its possible mechanisms[J]. Chin J Cancer Biother,2017,24(3):264-270. DOI:10.3872/j.issn.1007-385X.2017.03.009.
2019, Vol. 28 
