切换至 "中华医学电子期刊资源库"
高级检索
中华肺部疾病杂志(电子版)

中华肺部疾病杂志(电子版) ›› 2019, Vol. 12 ›› Issue (01) : 43 -48. doi: 10.3877/cma.j.issn.1674-6902.2019.01.008

所属专题： 文献；

论著

Rab26对小细胞肺癌H446细胞增殖和迁移的影响
陈伟1, 李华1, 唐心蔚1,†(), 刘雪萍1,†()   
  1. 1. 400037　重庆，陆军军医大学（第三军医大学）新桥医院胸外科
  • 收稿日期:2018-09-17 出版日期:2019-02-20
  • 通信作者: 唐心蔚, 刘雪萍
  • 基金资助:
    国家自然科学基金面上项目(81472188)

Effects of Rab26 on the proliferation and migration of small cell lung cancer cells H446

Wei Chen1, Hua Li1, Xinwei Tang1,†(), Xueping Liu1,†()   

  1. 1. Department of Chest surgery, Xinqiao Hospital, Army Medical University, Chongqing 400037, China
  • Received:2018-09-17 Published:2019-02-20
  • Corresponding author: Xinwei Tang, Xueping Liu
  • About author:
    Corresponding author: Liu Xueping, Email: ;
    Tang Xinwei,
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

陈伟, 李华, 唐心蔚, 刘雪萍. Rab26对小细胞肺癌H446细胞增殖和迁移的影响[J]. 中华肺部疾病杂志(电子版), 2019, 12(01): 43-48.

Wei Chen, Hua Li, Xinwei Tang, Xueping Liu. Effects of Rab26 on the proliferation and migration of small cell lung cancer cells H446[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2019, 12(01): 43-48.

目的

探讨Rab26对小细胞肺癌H446细胞增殖和迁移的影响。

方法

设立Rab26质粒过表达组及Rab26 siRNA组，培养H446细胞，分别将含有Rab26过表达质粒和Rab26 siRNA瞬时转染H446细胞，同时设立空白对照组。48 h后，流式细胞仪检测转染效率，同时检测每组细胞中Rab26在mRNA及蛋白水平表达情况；MTT检测每组细胞的增殖情况，划痕试验观察每组细胞迁移的速度。

结果

Rab26过表达质粒组转染效率为(76.8±4.3)%，Rab26 siRNA组转染效率为(79.5±3.57)%，均为有效转染；转染48 h后，发现Rab26质粒过表达组中Rab26在mRNA和蛋白水平均较空白对照组表达水平显著上调(P<0.05)，siRNA组结果显示Rab26 mRNA及蛋白表达水平显著下调(P<0.05)；MTT检测结果显示转染24 h和48 h后，空白对照组细胞活力分别为(56.4±3.23)%、(100±4.31)%；Rab26质粒过表达组细胞活力分别为(42.5±3.59)%、(62.3±2.97)%；Rab26 siRNA组细胞活力分别为(75±3.86)%、(123.4±5.66)%，即Rab26质粒过表达组细胞活力较空白组显著降低(P<0.05)，而Rab26 siRNA组细胞活力较空白组显著升高(P<0.05)，Rab26可抑制H446细胞的增殖；划痕实验结果显示转染24 h和48 h后，空白对照组细胞迁移率分别为(0.53±0.03)μm/min、(0.32±0.04)μm /min；Rab26质粒过表达组细胞迁移率分别为(0.21±0.04)μm/min、(0.22±0.04)μm/min；Rab26 siRNA组细胞迁移率分别为(0.61±0.02)μm/min、(0.42±0.03)μm/min，即Rab26质粒过表达组细胞迁移率较空白组显著较少(P<0.05)，而Rab26 siRNA组细胞迁移率较空白组显著增加(P<0.05)，Rab26可抑制H446细胞的迁移。

结论

Rab26可抑制小细胞肺癌H446细胞的增殖和迁移，故调控Rab26的表达有望成为小细胞肺癌治疗的新靶点。

关键词: Rab26, 小细胞肺癌, H446, 增殖, 迁移
Objective

To explore the effect of Rab26 on the Proliferation and migration of human in small cell lung cancer cells(H446).

Methods

The H446 cells were transfected with over-expression vector of Rab26, or Rab26 siRNA. Blank group was used as a negative control, respectively. After transfection 48 h, the protein level and mRNA of Rab26 were determined by RT-PCR and Western blot. Proliferation of the H446 cells was assayed by flow cytometry and the migration rate was detected by the scratch test.

Results

The transfection efficiency of Rab26 overexpression plasmid group was (76.8±4.3) % and Rab26 siRNA group was (79.5±3.57)%. After transfection 48 hours, the expression of Rab26 in overexpression group was significantly higher than control group in mRNA and protein level (P<0.05), however, the Rab siRNA group showed that the expression of Rab26 was significantly lower than control group (P<0.05). MTT assay showed that the cell viability of blank control group was (56.4±3.23)%, (100±4.31)%, Rab26 overexpression group was (42.5±3.59)%, (62.3±2.97)% and Rab26 siRNA group was (75±3.86)%, (123.31)% in 24 and 48 hours, respectively. The cell viability of Rab26 overexpression group was significantly lower than that of blank group (P<0.05), while Rab26 siRNA group was significantly higher than that of blank group (P<0.05). Scratch test showed that the cell migration rate of blank control group was (0.53±0.03)μm/min and (0.32±0.04)μm/min, Rab26 plasmid overexpression group was (0.21±0.04)μm/min and (0.22±0.04)μm/min, the Rab26 siRNA group was (0.61±0.02) μm/min, (0.42±0.03) um/min after 24 and 48 hours. The cell migration rates of Rab26 plasmid overexpression group was significantly lower than the blank group, but Rab26 siRNA group was significantly higher than the blank group (P<0.05).

Conclusions

Rab26 is associated with the cellular proliferation and migration in H446 cell. Rab26 may be a novel therapy target for the treatments of small cell lung cancer.

Key words: Rab26, Small cell lung cancer, H446 cells, Proliferation, Migration
图1 各组转染H446细胞的转染效率
图2 各组间Rab26mRNA和蛋白表达；注：*：P<0.05，与正常组比较
图3 各组细胞存活率；注：*: P<0.05，与对照24 h比较；#：P<0.05，与对照48 h比较
图4 各组细胞迁移情况
1
钱桂生. 肺癌不同病理类型发病率的变化情况及原因[J/CD]. 中华肺部疾病杂志(电子版), 2011, 4(1): 1-6.
2
胡正国，庞德湘. 中医治疗肺癌的研究进展[J]. 广西中医药大学学报，2015, 18(2): 84-86.
3
邹龑，李佩，唐卓. 小细胞肺癌诊断与化疗药物组合研究进展[J]. 四川医学，2011, 32(8): 1302-1304.
4
Fernandez-Espartero Ch,Ramel D,Farago M, et al. GTP exchange factor Vav regulates guided cell migration by coupling guidance receptor signalling to local Rac activation[J]. J Cell Sci, 2013, 126(10): 2285-2293.
5
陈华萍，陈旭昕，董伟杰，等. Rab26诱导宫颈癌Hela细胞凋亡并抑制其迁移的实验研究[J]. 第三军医大学学报，2016, 38(24): 1235-1238.
6
程颖. 小细胞肺癌治疗研究进展[J]. 中国处方药，2010, 101(8): 44- 47.
7
Johnson DH. Treatment of relapsed small cell lung cancer[J]. Lung Cancer, 1994, 11(Suppl 1): 142-143.
8
王金娜，王锦光，赵磊. 凋亡抑制基因Livin对肺腺癌细胞A549增殖与耐药的作用[J]. 中国医科大学学报，2015, 44(2): 114-118.
9
Baker S,Dahele M,Lagerwaard FJ, et al. A critical review of recent developments in radiotherapy for non-small cell lung cancer[J]. Radiat Oncol, 2016, 11(1): 115.
10
Paleiron N,Bylicki O,Andre M, et al. Targeted therapy for localized non-small-cell lung cancer: a review[J]. Onco Targets Ther, 2016, 9: 4099- 4104.
11
Counago F,Rodriguez A,Calvo P, et al. Targeted therapy combined with radiotherapy in non-small-cell lung cancer: a review of the Oncologic Group for the Study of Lung Cancer (Spanish Radiation Oncology Society)[J]. Clin Transl Oncol, 2017, 19(1): 31-43.
12
Yoshie S,Imai A,Nashida T, et al. Expression, characterization, and localization of Rab26, a low molecular weight GTP-binding protein, in the rat parotid gland[J]. Histochem Cell Biol, 2000, 113(4): 259-263.
13
Wagner AC,Strowski MZ,Goke B, et al. Molecular cloning of a new member of the Rab protein family, Rab 26, from rat pancreas[J]. Biochem Biophys Res Commun, 1995, 207(3): 950-956.
14
Nashida T,Imai A,Shimomura H. Relation of Rab26 to the amylase release from rat parotid acinar cells[J]. Arch Oral Biol, 2006, 51(2): 89-95.
15
Lo HG,Jin RU,Sibbel G, et al. A single transcription factor is sufficient to induce and maintain secretory cell architecture[J]. Genes Dev, 2017, 31(2): 154-171.
16
Jin RU,Mills JC. RAB26 coordinates lysosome traffic and mitochondrial localization[J]. J Cell Sci, 2014, 127(Pt 5): 1018-1032.
17
Mills JC,Taghert PH. Scaling factors: transcription factors regulating subcellular domains[J]. Bioessays, 2012, 34(1): 10-16.
18
Tian X,Jin RU,Bredemeyer AJ, et al. RAB26 and RAB3D are direct transcriptional targets of MIST1 that regulate exocrine granule maturation[J]. Mol Cell Biol, 2010, 30(5): 1269-1284.
19
Dong W,He B,Qian H, et al. RAB26-dependent autophagy protects adherens junctional integrity in acute lung injury[J]. Autophagy, 2018, 14(10): 1677-1692.
20
Li H,He B,Liu X, et al. Regulation on Toll-like Receptor 4 and Cell Barrier Function by Rab26 siRNA-loaded DNA Nanovector in Pulmonary Microvascular Endothelial Cells[J]. Theranostics, 2017, 7(9): 2537-2554.
21
Binotti B,Pavlos NJ,Riedel D, et al. The GTPase Rab26 links synaptic vesicles to the autophagy pathway[J]. Elife, 2015, 4: e05597.
22
Lo S,Yuan SS,Hsu C, et al. Lc3 over-expression improves survival and attenuates lung injury through increasing autophagosomal clearance in septic mice[J]. Ann Surg, 2013, 257(2): 352-363.
23
Barbieri MA,Roberts RL,Gumusboga A, et al. Epidermal growth factor and membrane trafficking：EGF receptor activation of endocytosis requires RAB5A[J]. J Cell Biol, 2000, 151(3): 539-550.
24
刘芳莉，李钰，高凌寒，等. RABSA基因表达改变对人肺腺癌细胞系GLC-82和SPC-al的分化和侵袭特性影响[J]. 遗传学报，2002, 29(12): 1043-1047.
25
史忠诚，于呖，李钰，等. RAB5A基因对肺腺癌细胞微丝的影响[J]. 遗传学报，2005, 32(12): 1227-1234.
26
Chen T,Han Y,Yang M, et al. Rab39, a novel Golgi-associated Rab GTPase from human dendriticcells involved in cellular endocy tosis[J]. Biochem Biophys Res Commo, 2003, 303(4): 1114-1120.
27
Mai A,Veltel S,Pellinen T, et al. Competitive binding of Rab21 and p120RasGAP to integrins regulates receptor traffic and migration[J]. J Cell Biol, 2011, 194(2): 291-306.
28
Cao C,Lu C,Xu J, et al. Expression of Rab25 correlates with the invasion and metastasis of gastric cancer[J]. Chin J Cancer Res, 2013, 25(2): 192-199.
29
Zilber Y,Babayeva S,Seo JH, et al. The PCP effector Fuzzy controls cilial assembly and signaling by recruiting Rab8 and disheveled to the primary cilium[J]. Mol Biol Cell, 2013, 24(5): 555-565.
30
张万秋，田寅灰，朱一超. Rab基因家族在MDA-MB-231乳腺癌细胞胞体和伪足中的差异表达[J]. 南京医科大学学报，2012, 32(9): 1192-1198.
31
Mili S,Moissoglu K,Macara G. Genome wide screen reveals APC-associated RNAs enriched in cell protrusions[J]. Nature, 2008, 453(7191): 115-119.
32
Mukherjee S,Liu X,Arasaki K, et al. Roy CR. Modulation of Rab GTPase function by a protein phosphocholine transferase[J]. Nature, 2011, 477(7362): 103-106.
[1] 黄福, 王黔, 金相任, 唐云川. VEGFR2、miR-27a-5p在胃癌组织中的表达与临床病理参数及预后的关系研究[J]. 中华普外科手术学杂志(电子版), 2024, 18(05): 558-561.
[2] 胡思平, 熊性宇, 徐航, 杨璐. 衰老相关分泌表型因子在前列腺癌发生发展中的作用机制[J]. 中华腔镜泌尿外科杂志(电子版), 2024, 18(05): 425-434.
[3] 赵蒙蒙, 黄洁, 余荣环, 王葆青. 过表达小GTP酶Rab32抑制非小细胞肺癌细胞侵袭性生长[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 512-518.
[4] 张桂萍, 丘勇林, 湛绮婷, 孙乐栋. 晚期非小细胞肺癌血清Ape1/Ref-1对放射性肺损伤发生的预测意义[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 519-523.
[5] 韩晓宇, 李柯育, 赵志菲, 高建平. SNHG17过表达对非小细胞肺癌切除术预后的意义[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 543-547.
[6] 刘松, 张进召, 贾艳云. 帕博利珠单抗治疗晚期非小细胞肺癌反应降低与抗生素预处理的关系[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 553-557.
[7] 李多, 郝昭昭, 陈延伟, 南岩东. 血清PTX3表达与非小细胞肺癌骨转移的相关性分析[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 558-562.
[8] 曾聿理, 雷发容, 肖慧, 邱德亮, 谢静, 吴寻. 氯普鲁卡因通过调控circRNA-ZKSCAN1表达抑制肝癌Huh-7细胞体外生长和转移的研究[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 220-228.
[9] 李彦浇, 梁雷, 金钫, 王智伟. 银杏内酯B通过调控miR-24-3p对人牙周膜干细胞增殖、成骨分化的影响[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 229-235.
[10] 李晶, 潘侠, 周芳, 汪晶, 洪佳. 普鲁卡因通过上调lncRNA DGCR5抑制胃癌细胞增殖、迁移和侵袭[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 151-158.
[11] 刘杜先, 张杰东, 付鲁渝, 熊志强, 龚程, 张小雅, 高明悦, 孟俊宏, 刘兰侠. 沉默circXPO1抑制肝癌细胞恶性生物学行为[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 159-166.
[12] 李博, 马秀岩, 孙杰. lncRNA TINCR对滋养层HTR-8/SVneo细胞生物学行为的影响及其机制[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 167-172.
[13] 崔精, 鲍一帆, 沈晓明, 杨增辉, 高森, 鲍传庆. 结直肠癌中circMFSD12对肿瘤细胞功能及5-FU敏感性的调控[J]. 中华结直肠疾病电子杂志, 2024, 13(04): 294-302.
[14] 靳英, 付小霞, 陈美茹, 袁璐, 郝力瑶. CD147调控MAPK信号通路对结肠癌细胞增殖和凋亡的影响及机制研究[J]. 中华临床医师杂志(电子版), 2024, 18(05): 474-480.
[15] 张迅夫, 马金山, 蒋云龙, 加纳提·托勒恒, 侯昌剑, 萨伍提·斯拉吉丁. GATA3在非小细胞肺癌组织中的表达及临床病理意义[J]. 中华胸部外科电子杂志, 2024, 11(03): 175-179.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号