中国癌症杂志 ›› 2014, Vol. 24 ›› Issue (7): 529-534.doi: 10.3969/j.issn.1007-3969.2014.07.010

• 论著 • 上一篇    下一篇

下调肌动蛋白相关蛋白2/3复合体5对肺鳞癌细胞株SK-MES-1增殖、侵袭和转移能力的影响

杜江,张林   

  1. 中国医科大学附属一院胸外科,辽宁 沈阳 110001
  • 出版日期:2014-07-30 发布日期:2014-07-29
  • 通信作者: 张林 E-mail:zhanglin_cmu@126.com

Downregulation of actin related protein 2/3 complex subunit 5 inhibits proliferation, migration and invasion of lung squamous carcinoma cell line SK-MES-1

DU Jiang,ZHANG Lin   

  1. Department of Thoracic Surgery, China Medical University Affiliated No.1 Hospital, Shenyang Liaoning 110001, China
  • Published:2014-07-30 Online:2014-07-29
  • Contact: ZHANG Lin E-mail: zhanglin_cmu@126.com

摘要:

背景与目的:肌动蛋白相关蛋白2/3复合体5(actin related protein 2/3 complex subunit 5ARPC5)参与肌动蛋白的组装,影响细胞的运动能力,在形成癌细胞侵袭性伪足的过程中起到重要作用。本研究通过基因沉默手段,探讨下调ARPC5基因表达对肺鳞癌细胞株SK-MES-1增殖、侵袭和转移能力的影响。方法:应用阳离子脂质体转染法构建ARPC5基因沉默(si-ARPC5)SK-MES-1肺鳞癌细胞株,qRT-PCR和蛋白质印迹法(Western blot)鉴定转染效率和表达情况。实验设置si-ARPC5实验组、空白对照组(mock)及阴性对照组(NC)。采用细胞技术试剂盒(cell couiting kit-8CCK-8)法测定si-ARPC5SK-MES-1细胞的增殖的影响;划痕修复实验测定si-ARPC5SK-MES-1细胞迁移能力的影响;Transwell小室体外侵袭实验测定si-ARPC5SK-MES-1细胞的侵袭能力的影响。结果:转染si-ARPC5后的SK-MES-1细胞在mRNA和蛋白水平上均下调ARPC5的表达。与mock组及NC组比较,si-ARPC5实验组SK-MES-1细胞增殖能力明显下降(t=7.993t=8.681P<0.05)si-ARPC5实验组划痕修复率(43.32±0.23)%,而mock组和NC组划痕修复率分别为(73.11±0.43)%(76.58±0.88)%,差异有统计学意义(t=7.348t=10.614P<0.05)si-ARPC5组的SK-MES-1细胞穿膜数为(27±6)个,mock组为(101±11)个,NC组为(92±9)个。si-ARPC5组穿膜细胞数明显少于其他2组,差异有统计学意义(t=10.229t=8.391P<0.05)结论:下调ARPC5基因的表达能够显著抑制肺鳞癌细胞株 SK-MES-1的增殖、侵袭和转移能力。

关键词: 肺鳞癌, 肌动蛋白相关蛋白2/3复合体5, 增殖, 迁移, 侵袭

Abstract:

Background and purpose: Actin related protein 2/3 complex subunit 5 (ARPC5) is involved in the packaging of actin, and then affects the mobility of cells. It plays an important role in the formation of invadopodia of cancer cells. This study aimed to investigate the effect of ARPC5 gene-silence on cell proliferation, migration and invasion of lung squamous carcinoma cell line SK-MES-1. Methods: Under the induction of LipofectamineTM RNAiMAX, the recombinant of si-ARPC5 was transfected into SK-MES-1. The experiment set up 3 groups: blank (mock) group, negative control (NC) group and si-ARPC5 transfected group. Cell proliferation, migration and invasion assays were investigated by cell counting kit(CCK-8) method, scratch assay and Transwell Chambers respectively. Results: The result of qRT-PCR and Western blot displayed downregulation of ARPC5 in mRNA and protein level of SK-MES-1 cell. Proliferation rate of SK-MES-1 cells in 72 h after si-ARPC5 transfection was lower than those in mock and NC groups (t=7.993, t=8.681, P<0.05). Compared to the mock and NC groups, the scratch repairing rate of the si-ARPC5 group was lower [(43.32±0.23)% vs (73.11±0.43)% and (76.58±0.88)%] (t=7.348, t=10.614, P<0.05); And the number of penetrating the membrane was significantly reduced [(27±6) vs (101±11) and (92±9)] (t=10.229, t=8.391, P<0.05). Conclusion: Downregulation of ARPC5 gene can inhibit the proliferation, migration, and invasion of SKMES-1 cell.

Key words: Lung squamous cell carcinoma, Actin related protein 2/3 complex subunit 5, Proliferation, Migration, Invasion