小分子干扰RNA抑制HLX1表达对急性髓系白血病细胞侵袭的影响及机制探讨

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (6): 627-632.

小分子干扰RNA抑制HLX1表达对急性髓系白血病细胞侵袭的影响及机制探讨

沈健，林颖超，朱夏吟

台州医院血液肿瘤科，临海 317000，浙江

收稿日期:2017-01-24 修回日期:2017-04-14 出版日期:2017-06-26 发布日期:2017-06-26
作者简介:沈健，男，硕士研究生，主治医生，研究方向：淋巴瘤临床治疗。 Tel：13758628257 E-mail:hyper1001@126.com
基金资助:
浙江省医药卫生项目（2017195761）

Inhibitory effect of HLX1 on proliferation and invasion of acute myeloid leukemia cell

SHEN Jian, LIN Yingchao, ZHU Xiayin

Deparment of Hematology and Oncology, Taizhou Hospital, Linhai 317000, Zhejiang, China

Received:2017-01-24 Revised:2017-04-14 Online:2017-06-26 Published:2017-06-26

摘要/Abstract

摘要：

目的: 探讨小分子干扰RNA（siRNA）抑制人同源异型盒基因（H2.0-like homeobox，HLX1）表达对急性髓系白血病细胞系U937细胞增殖与侵袭能力的影响。方法: 利用HLX1特异的siRNA瞬时转染U937细胞，通过实时定量（RT-PCR）与蛋白印迹法（Western blot）分别测定HLX1的mRNA及蛋白质的表达水平，应用细胞体外增殖实验法（MTT）与体外侵袭实验观察siRNA抑制HLX1表达后的U937细胞增殖与侵袭变化。结果: 相对于空白对照组，HLX1特异的siRNA转染24 h后可抑制U937细胞HLX1 mRNA表达，抑制率为（70.0±2.7）%（P<0.01）；转染48 h后可有效抑制HLX1蛋白质表达，抑制率为（81.0±3.1）%（P<0.01）；转染组U937细胞增殖能力显著下降（P<0.01），转染组穿过基质胶的U937细胞数（22.0±2.3）显著低于空白对照组（66.0±5.0）(P<0.01)， p-21活化激酶1（PAK1）蛋白表达水平显著下降(P<0.01)。结论: siRNA下调HLX1表达水平可显著抑制急性髓系白血病细胞的增殖与侵袭能力。

关键词: 急性髓系白血病细胞, U937, 同源异型盒基因, RNA干扰, 侵袭

Abstract:

AIM: To explore the effect of HLX1 on the proliferation and invasion ability in acute myeloid leukemia cell U937. METHODS: The HLX1 mRNA-targeting siRNA was transfected to U937 cells, the mRNA and protein expression of HLX1 were assessed by RT-PCR and Western blot, respectively. MTT assay and Transwell chamber assay were used to observe the proliferation and invasion ability of U937 cell after being transfected with HLX1 mRNA-targeting siRNA. RESULTS:Compared with the control group, HLX1 siRNA transfection reduced the level of HLX1 mRNA in U937 cells after 24 h, the inhibition rate was (70.0±2.7)% (P<0.01)； HLX1 siRNA transfection also reduced the expression of HLX1 protein in U937 cells after 48 h, the inhibition rate was (81.0±3.1)% (P<0.01); HLX1 transfection group cell proliferation was inhibited significantly (P<0.01); The invasion cells of HLX1 siRNA(22.0±2.3) were significantly lower than that of control group(66.0±5.0)(P<0.01), the expression of PAK1 protein was decreased in HLX1 transfection group(P<0.01). CONCLUSION: SiRNA can significantly inhibit the proliferation and invasion of acute myeloid leukemia cell by down-regulating the expression of HLX1.

Key words: acute myeloid leukemia cell, U937, HLX1, RNA interference, invasion

中图分类号:

R965.2
R552

沈健，林颖超，朱夏吟. 小分子干扰RNA抑制HLX1表达对急性髓系白血病细胞侵袭的影响及机制探讨[J]. 中国临床药理学与治疗学, 2017, 22(6): 627-632.

SHEN Jian, LIN Yingchao, ZHU Xiayin. Inhibitory effect of HLX1 on proliferation and invasion of acute myeloid leukemia cell[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(6): 627-632.

参考文献

［1］Cardarelli F, Bijol V, Chandraker A, et al. Acute myeloid leukemia after kidney transplantation: a case report and literature review［J］. J Bras Nefrol, 2016, 38(4):455-461.
［2］Meyer SC, Levine RL. Translational implications of somatic genomics in acute myeloid leukaemia［J］. Lancet Oncol, 2014, 15(9): e382-e394.
［3］Estey E. Acute myeloid leukemia-many diseases, many treatments［J］. N Engl J Med, 2016, 375(21): 2094-2095.
［4］Chen X, Xie H, Wood BL, et al. Relation of clinical response and minimal residual disease and their prognostic impact on outcome in acute myeloid leukemia［J］. J Clin Oncol, 2015, 33(11):1258-1264.
［5］Liu H, Murthi P, Qin S, et al. A novel combination of homeobox genes is expressed in mesenchymal chorionic stem/stromal cells in first trimester and term pregnancies［J］. Reprod Sci, 2014, 21(11):1382-1394.
［6］Chinen Y, Taki T, Tsutsumi Y, et al. The leucine twenty homeobox (LEUTX) gene, which lacks a histone acetyltransferase domain, is fused to KAT6A in therapy-related acute myeloid leukemia with t(8;19)(p11;q13)［J］. Genes Chromosomes Cancer, 2014, 53(4):299-308.
［7］Abedin S, Altman JK. Acute promyelocytic leukemia: preventing early complications and late toxicities［J］. Hematol Am Soc Hematol Educ Program, 2016, 1:10-15.
［8］Ito Y. Treatment and clinicopathological features of relapsed, refractory or elderly AML［J］. Rinsho Ketsueki, 2015, 56(10):1960-1968.
［9］Mi JQ, Chen SJ, Zhou GB, et al. Synergistic targeted therapy for acute promyelocytic leukaemia: a model of translational research in human cancer［J］. J Intern Med, 2015, 278(6):627-642.
［10］De K, Abdulhay M. Acute myeloid leukemia: a comprehensive review and 2016 update［J］. Blood Cancer Journal, 2016, 6: e441.
［11］Xu Y, Gao J, Su Z, et al. Downregulation of Hlx closely related to the decreased expressions of T-bet and Runx3 in patients with gastric cancer may be associated with a pathological event leading to the imbalance of Th1/Th2［J］. Clin Dev Immunol, 2012, 2012: 949821.
［12］Liu T, Chen J, Xiao S, et al. H2.0-like homeobox 1 acts as a tumor suppressor in hepatocellular carcinoma［J］. Tumor Biol, 2016, 37(5):6419-6428.
［13］Shields BJ, Jackson JT, Metcalf D, et al. Acute myeloid leukemia requires Hhex to enable PRC2-mediated epigenetic repression of Cdkn2a［J］. Genes Dev, 2016, 30(1):78-91.
［14］Kawahara M, Pandolfi A, Bartholdy B, et al. H2.0-like homeobox regulates early hematopoiesis and promotes acute myeloid leukemia［J］. Cancer Cell, 2012, 22(2):194-208.
［15］Vukovic M, Guitart AV, Sepulveda C, et al. Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance ［J］. J Exp Med, 2015, 212(13):2223-2234.
［16］Zhan MN, Yu XT, Tang J, et al. MicroRNA-494 inhibits breast cancer progression by directly targeting PAK1［J］. Cell Death Dis, 2017, 8(1): e2529.
［17］Ong CC, Jubb AM, Zhou W, et al. p21-activated kinase 1: PAK'ed with potential［J］. Oncotarget, 2011, 2(6): 491-496.

[1]	王艳, 张金辉, 赵永辰, 刘宏祥, 刘亚维, 苗欢欢, 杨信才. 红杉黄酮通过下调PI3K/AKT信号通路抑制胃癌细胞增殖侵袭等干细胞特性[J]. 中国临床药理学与治疗学, 2023, 28(5): 508-513.
[2]	眭玉霞, 庄捷, 庄敏, 黄桂. Delicaflavone通过PI3K/AKT/mTOR通路调控上皮-间质转化抑制吉非替尼耐药肺癌PC-9/GR细胞迁移和侵袭[J]. 中国临床药理学与治疗学, 2022, 27(6): 614-621.
[3]	施经斌, 熊阳. 肿瘤微环境与乳腺癌细胞互作导致肿瘤转移的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(5): 562-574.
[4]	李岩岩, 方骁杰, 尹鑫, 孙曦, 饶春晖. circZNF124通过靶向miR-4262调控结直肠癌SW620细胞增殖、迁移及侵袭的机制研究[J]. 中国临床药理学与治疗学, 2022, 27(11): 1231-1239.
[5]	侯彬, 黄维平, 尹中普, 胡守森. lncRNA PCAT19吸附miR-142-5p调控ING3基因表达抑制鼻咽癌细胞的增殖和侵袭[J]. 中国临床药理学与治疗学, 2020, 25(7): 721-727.
[6]	沈成，杨年钊，王明海，吴立胜，朱志强. RNA甲基化酶NSUN2对胃癌细胞侵袭和迁移的影响及机制研究[J]. 中国临床药理学与治疗学, 2020, 25(4): 380-386.
[7]	张康，何斌军，胡汶斌，韩晓亮. 碳酸酐酶1沉默对肺癌A549细胞增殖、凋亡、侵袭和迁移的影响[J]. 中国临床药理学与治疗学, 2020, 25(1): 55-60.
[8]	王庆玮，丁锦，吕爰爰，罗贤臣，倪观太. PHA-767491抑制CDC-7的表达在子宫内膜癌ishikawa细胞系中的作用研究[J]. 中国临床药理学与治疗学, 2019, 24(8): 866-871.
[9]	周青英，徐乃奋，郑定钦，潘其壮，张志刚，汪朔. 氟尿嘧啶调控VHL/HIF信号通路抑制肾癌细胞增殖、侵袭与转移的研究[J]. 中国临床药理学与治疗学, 2019, 24(8): 872-879.
[10]	朱林佳，原少斐，上官宗校，慈晓，赵仁国，李玉苹. microRNA-1304通过靶向血红素氧合酶-1对人肺癌细胞的抑制作用[J]. 中国临床药理学与治疗学, 2019, 24(4): 383-390.
[11]	陈悦群，翟洪波，仝进毅，屈王蕾. 竹节香附素通过抑制JAK/STAT3信号转导通路抑制人子宫内膜癌HEC-1-B细胞的侵袭转移[J]. 中国临床药理学与治疗学, 2019, 24(4): 391-396.
[12]	李鹏，杨荣华，张明华，肖鑫，汤建儿. miR-34a-5p在膀胱癌组织中的表达及其对膀胱癌细胞侵袭迁移的影响和机制[J]. 中国临床药理学与治疗学, 2019, 24(4): 403-410.
[13]	钱万锋，王秀峰，陈学鹏. miR-19b在骨肉瘤组织中的表达及其通过调控KLF13影响骨肉瘤Saos-2细胞侵袭和迁移的研究[J]. 中国临床药理学与治疗学, 2019, 24(11): 1242-1248.
[14]	赵巧素，叶桦，孙勤学. 生物钟基因Period2调节肝癌细胞warburg效应及转移侵袭中的作用机制[J]. 中国临床药理学与治疗学, 2019, 24(10): 1134-1141.
[15]	孙红亚，袁爱娟，李纪鹏. 胡桃醌对人卵巢癌SKOV3和IOSE80细胞增殖和侵袭的抑制作用及其机制研究[J]. 中国临床药理学与治疗学, 2019, 24(1): 32-37.