Effect of PTTG1 Expression on Invasion and Migration of Colon Cancer Cell SW480 and Its Possible Mechanism
-
摘要:目的
研究垂体肿瘤转化基因 1（pituitary tumor transforming gene 1,PTTG1）过表达促进人结肠癌细胞SW480侵袭和迁移作用及其可能机制。
方法采用脂质体转染法将pcDNA3.1(+)-PTTG1及空载pcDNA3.1(+)转染人结肠癌SW480细胞，G418法筛选阳性克隆。Western blot和Real-time PCR法鉴定稳定过表达PTTG1细胞株建立。Transwell小室法检测细胞侵袭和迁移能力，Western blot检测MMP2、MMP9、E-cadherin、Vimentin和Snail的表达。
结果（1）成功获得稳定高表达PTTG1的SW480克隆细胞株PTTG1-SW480；（2）过表达PTTG1基因后，SW480细胞侵袭和迁移能力增强，MMP2和MMP9表达升高，上皮间质转化（epithelial-mesenchymal transition,EMT）标记分子E-cadherin表达降低，Vimentin和Snail的表达升高，差异均有统计学意义（P＜0.01）；（3）过表达PTTG1基因后，SW480细胞中PI3K/AKT信号活化增强，使用LY29400干预后，抑制细胞侵袭、迁移和EMT，E-cadherin表达上调、Vimentin和Snail的表达下调，差异均有统计学意义（P＜0.01）。
结论PTTG1基因过表达可能通过活化PI3K/AKT信号诱导SW480细胞EMT发生，发挥促进SW480侵袭和迁移作用；提示PTTG1蛋白可能成为结肠癌治疗的一个潜在靶点。
Abstract:ObjectiveTo investigate the effect of over-expression of pituitary tumor transforming gene 1 (PTTG1) on the invasion and migration of human colon cancer cells SW480 and its possible mechanism.
MethodsBoth pcDNA3.1(+)-PTTG1and pcDNA3.1(+) plasmids were separately transfected into human colon cancer cell line SW480 by liposome transfection method and G418 was used to screen positive clones. Real-time PCR and Western blot were used to asses the stably over-expressed PTTG1 in PTTG1-SW480. The abilities of migration and invasion of PTTG1-SW480 cells were assayed by Transwell chambers and Western blot was used to detect the expression of MMP2,MMP9,E-cadherin,Vimentin and Snail.
Results(1) The cloned cells PTTG1-SW480 with PTTG1 over-expression were successfully obtained; (2) After PTTG1 over-expression,the abilities of migration and invasion of SW480 cells were enhanced,the expression of MMP2 and MMP9 were up-regulated,Vimentin and Snail expression were up-regulated,and E-cadherin expression was down-regulated (all P＜0.01) ; (3) The signaling pathway of PI3K/AKT activation was enhanced in PTTG1-SW480 cells. Interfering with LY29400,the abilities of migration and invasion and EMT of PTTG1-SW480 cells were prevented,the expression of E-cadherin was up-regulated and Vimentin and Snail expression were down-regulated (all P＜0.01) .
ConclusionPTTG1 over-expression may promote SW480 cells migration and invasion through activating PI3K/AKT pathway to induce EMT. PTTG1 may become a potential target for colon cancer treatment.
-
Key words:
- PTTG1 /
- Colon cancer /
- Invasion /
- Migration
-
-
![]()
图 3 PTTG1促进SW480细胞中MMP2和MMP9的表达
Figure 3 PTTG1 up-regulated MMP2 and MMP9 expression in SW480 cells
![]()
图 4 PTTG1促进SW480细胞上皮间质转化
Figure 4 PTTG1 promoted epithelial-mesenchymal transition(EMT) of SW480 cells
![]()
图 5 PTTG1促进SW480细胞PI3K/AKT信号活化
Figure 5 PTTG1 enhanced activation of signal pathway of PI3K/AKT in SW480 cells
![]()
图 6 PTTG1通过活化PI3K/AKT信号促进SW480细胞侵袭与迁移
Figure 6 PTTG1 promoted SW480 cells invasion and migration through activating signal pathway of PI3K/AKT
-
[1] Center MM,Jemal A,Smith RA,et al. Worldwide variations in colorectal cancer[J]. CA Cancer J Clin,2009,59(6): 366-78. doi: 10.3322/caac.20038
[1] Center MM, Jemal A, Smith RA, et al. Worldwide variations in colorectal cancer[J]. CA Cancer J Clin, 2009, 59(6): 366-78. [2] McMillan DC, McArdle CS. Epidemiology of colorectal liver metastases[J]. Surg Oncol, 2007, 16(1): 3-5. [2] McMillan DC,McArdle CS. Epidemiology of colorectal liver metastases[J]. Surg Oncol,2007,16(1): 3-5. doi: 10.1016/j.suronc.2007.04.008
[3] Cunningham D, Atkin W, Lenz HJ, et al. Colorectal cancer[J]. Lancet, 2010, 375(9719): 1030-47. [3] Cunningham D,Atkin W,Lenz HJ,et al. Colorectal cancer[J]. Lancet,2010,375(9719): 1030-47. doi: 10.1016/S0140-6736(10)60353-4
[4] Noll JE,Vandyke K,Hewett DR,et al. PTTG1 expression is associated with hyperproliferative disease and poor prognosis in multiple myeloma[J]. J Hematol Oncol,2015,8(1): 106. doi: 10.1186/s13045-015-0209-2
[4] Noll JE, Vandyke K, Hewett DR, et al. PTTG1 expression is associated with hyperproliferative disease and poor prognosis in multiple myeloma[J]. J Hematol Oncol, 2015, 8(1): 106. [5] Li L,Han LY,Yu M,et al. Pituitary tumor-transforming gene 1 enhances metastases of cervical cancer cells through miR-3666-regulated ZEB1[J]. Tumour Biol,2015,[Epub ahead of print]. http://cn.bing.com/academic/profile?id=1941520307&encoded=0&v=paper_preview&mkt=zh-cn
[5] Li L, Han LY, Yu M, et al. Pituitary tumor-transforming gene 1 enhances metastases of cervical cancer cells through miR-3666- regulated ZEB1[J]. Tumour Biol, 2015, [Epub ahead of print]. [6] Heaney AP,Singson R,McCabe CJ,et al. Expression of pituitary-tumour transforming gene in colorectal tumours[J]. Lancet,2000,355(9205): 716-9. doi: 10.1016/S0140-6736(99)10238-1
[6] Heaney AP, Singson R, McCabe CJ, et al. Expression of pituitarytumour transforming gene in colorectal tumours[J]. Lancet, 2000, 355(9205): 716-9. [7] Lin YH, Tian Y, Wang JS, et al. Pituitary tumor-transforming gene 1 regulates invasion of prostate cancer cells through MMP13[J]. Tumour Biol, 2015, [Epub ahead of print]. [7] Lin YH,Tian Y,Wang JS,et al. Pituitary tumor-transforming gene 1 regulates invasion of prostate cancer cells through MMP13[J]. Tumour Biol,2015,[Epub ahead of print]. http://cn.bing.com/academic/profile?id=1029209396&encoded=0&v=paper_preview&mkt=zh-cn
[8] Zuo QF,Cao LY,Yu T,et al. MicroRNA-22 inhibits tumor growth and metastasis in gastric cancer by directly targeting MMP14 and snail[J]. Cell Death Dis,2015,6: e2000. doi: 10.1038/cddis.2015.297
[8] Zuo QF, Cao LY, Yu T, et al. MicroRNA-22 inhibits tumor growth and metastasis in gastric cancer by directly targeting MMP14 and snail[J]. Cell Death Dis, 2015, 6: e2000. [9] Kumar K,Chow CR,Ebine K,et al. Differential Regulation of ZEB1 and EMT by MAPK-interacting protein kinases (MNKs) and eIF4E in Pancreatic Cancer[J]. Mol Cancer Res,2015,14(2): 216-27. http://cn.bing.com/academic/profile?id=2173736103&encoded=0&v=paper_preview&mkt=zh-cn
[9] Kumar K, Chow CR, Ebine K, et al. Differential Regulation of ZEB1 and EMT by MAPK-interacting protein kinases (MNKs) and eIF4E in Pancreatic Cancer[J]. Mol Cancer Res, 2015, 14(2): 216-27. [10] Zhu QQ, Ma C, Wang Q, et al. The role of TWIST1 in epithelialmesenchymal transition and cancers[J]. Tumour Biol, 2016, 37(1): 185-97. [10] Zhu QQ,Ma C,Wang Q,et al. The role of TWIST1 in epithelial-mesenchymal transition and cancers[J]. Tumour Biol,2016,37(1): 185-97. doi: 10.1007/s13277-015-4450-7
[11] Yoon CH, Kim MJ, Lee H, et al. PTTG1 oncogene promotes tumor malignancy via epithelial to mesenchymal transition and expansion of cancer stem cell population[J]. J Biol Chem, 2012, 287(23): 19516-27. [11] Yoon CH,Kim MJ,Lee H,et al. PTTG1 oncogene promotes tumor malignancy via epithelial to mesenchymal transition and expansion of cancer stem cell population[J]. J Biol Chem,2012,287(23): 19516-27. doi: 10.1074/jbc.M111.337428
[12] Zhang E, Liu S, Xu Z, et al. Pituitary tumor-transforming gene 1 (PTTG1) is overexpressed in oral squamous cell carcinoma (OSCC) and promotes migration, invasion and epithelialmesenchymal transition (EMT) in SCC15 cells[J]. Tumour Biol, 2014, 35(9): 8801-11. [12] Zhang E,Liu S,Xu Z,et al. Pituitary tumor-transforming gene 1 (PTTG1) is overexpressed in oral squamous cell carcinoma (OSCC) and promotes migration,invasion and epithelial-mesenchymal transition (EMT) in SCC15 cells[J]. Tumour Biol,2014,35(9): 8801-11. doi: 10.1007/s13277-014-2143-2
[13] Brzozowa M,Michalski M,Wyrobiec G,et al. The role of Snail1 transcription factor in colorectal cancer progression and metastasis[J]. Contemp Oncol(Pozn),2015,19(4): 265-70. http://cn.bing.com/academic/profile?id=2413329656&encoded=0&v=paper_preview&mkt=zh-cn
[13] Brzozowa M, Michalski M, Wyrobiec G, et al. The role of Snail1 transcription factor in colorectal cancer progression and metastasis[J]. Contemp Oncol(Pozn), 2015, 19(4): 265-70. [14] Wang SC, Chai DS, Chen CB, et al. HPIP promotes thyroid cancer cell growth, migration and EMT through activating PI3K/AKT signaling pathway[J]. Biomed Pharmacother, 2015, 75: 33-9. [14] Wang SC,Chai DS,Chen CB,et al. HPIP promotes thyroid cancer cell growth,migration and EMT through activating PI3K/AKT signaling pathway[J]. Biomed Pharmacother,2015,75: 33-9. doi: 10.1016/j.biopha.2015.08.027
[15] Chen X,Liao J,Lu Y,et al. Activation of the PI3K/Akt pathway mediates bone morphogenetic protein 2-induced invasion of pancreatic cancer cells Panc-1[J]. Pathol Oncol Res,2011,17(2): 257-61. doi: 10.1007/s12253-010-9307-1
[15] Chen X, Liao J, Lu Y, et al. Activation of the PI3K/Akt pathway mediates bone morphogenetic protein 2-induced invasion of pancreatic cancer cells Panc-1[J]. Pathol Oncol Res, 2011, 17(2): 257-61. [16] Wang F, Ruan XJ, Zhang HY. BDE-99 (2,2',4,4',5-pentabromodiph enyl ether) triggers epithelial-mesenchymal transition in colorectal cancer cells via PI3K/Akt/Snail signaling pathway[J]. Tumori, 2015, 101(2): 238-45. [16] Wang F,Ruan XJ,Zhang HY. BDE-99 (2,2',4,4',5-pentabromodiphenyl ether) triggers epithelial-mesenchymal transition in colorectal cancer cells via PI3K/Akt/Snail signaling pathway[J]. Tumori,2015,101(2): 238-45. doi: 10.5301/tj.5000229
[17] Cho SH,Park YS,Kim HJ,et al. CD44 enhances the epithelial-mesenchymal transition in association with colon cancer invasion[J]. Int J Oncol,2012,41(1): 211-8.
[17] Cho SH, Park YS, Kim HJ, et al. CD44 enhances the epithelialmesenchymal transition in association with colon cancer invasion[J]. Int J Oncol, 2012, 41(1): 211-8.
下载:
计量
- 文章访问数:
- HTML全文浏览量: 0
- PDF下载量:
