Effect of PTEN siRNA on Proliferation and Invasion in Colon Cancer Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 LIYuan ¹ ,  MAJia-chi ¹ , LIYi-ping ¹ , FANGWei ¹ , GUOQing-jin ²

1. Department of General Surgery, Gansu Provincial People's Hospital, Lanzhou 730000, Gansu Province, China;
2. Graduate School, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China;

Corresponding author：

MAJia-chi, Email: tsmjc@hotmail.com

Keywords：

PTEN; siRNA; Colon cancer; Proliferation; Invasion

DOI：

10.7507/1007-9424.20150145

Video：

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Objective To explore the influence mechanism of proliferation and invasion in colon cancer cell after silence of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene. Methods RT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and Colo320 cell lines). small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WsT-1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells. Results PTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P < 0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P < 0.01). Conclusions PTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.

Citation： LIYuan, MAJia-chi, LIYi-ping, FANGWei, GUOQing-jin. Effect of PTEN siRNA on Proliferation and Invasion in Colon Cancer Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2015, 22(5): 544-548. doi: 10.7507/1007-9424.20150145 Copy

1.	Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin, 2011, 61(2):69-90.
2.	shaukat A, Mongin sJ, Geisser Ms, et al. long-Term mortality after screening for colorectal cancer. N Engl J Med, 2013, 369(12):1106-1114.
3.	汪晓东, 李立.结直肠癌外科的新理念与挑战.中国普外基础与临床杂志, 2014, 21(6):782-786.
4.	siegel R, Desantis C, Jemal A. Colorectal cancer statistics, 2014. CA Cancer J Clin, 2014, 64(2):104-117.
5.	stambolic V, suzuki A, De la Pompa Jl, et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell, 1998, 95(1):29-39.
6.	Ma JC, sawai H, Matsuo Y, et al. Interleukin-1alpha enhances angiogenesis and is associated with liver metastatic potential in human gastric cancer cell lines. J Surg Res, 2008, 148(2):197-204.
7.	Ma JC, sawai H, Matsuo Y, et al. IGF-1 mediates PTEN suppression and enhances cell invasion and proliferation via activation of the IGF-1/PI3K/Akt signaling pathway in pancreatic cancer cells. J Surg Res, 2010, 160(1):90-101.
8.	张永红, 于冬冬, 何乐亚, 等.结肠癌原发灶及肝转移灶中PTEN蛋白的表达研究.中国组织化学与细胞化学杂志, 2010, 19(1):85-87.
9.	艾源.结直肠癌患者是否存在PTEN基因突变?中国普外基础与临床杂志, 2013, 20(7):811.
10.	Hollander MC, Blumenthal GM, Dennis PA. PTEN loss in the continuum of common cancers, rare syndromes and mouse models. Nat Rev Cancer, 2011, 11(4):289-301.
11.	刘丹, 徐建洪, 焦泽龙, 等. PTEN基因功能及其调控研究进展.解放军医药杂志, 2014, 26(4):111-116.
12.	Ma JC, sawai H, Matsuo Y, et al. IGF-1 and PTEN regulate the proliferation and invasiveness of colon cancer cells through opposite effects on PI3K/Akt signalling. Arch Med Sci, 2009, 2:195-206.
13.	Ma JC, sawai H, Ochi N, et al. PTEN regulates angiogenesis through PI3K/Akt/VEGF signaling pathway in human pancreatic cancer cells. Mol Cell Biochem, 2009, 331(1-2):161-171.
14.	Hempel N, Melendez JA. Intracellular redox status controls membrane localization of pro-and anti-migratory signaling molecules. Redox Biol, 2014, 20(2):245-250.
15.	Xu BF, Washington AM, Hinton BT. PTEN signaling through RAF1 proto-oncogene serine/threonine kinase (RAF1)/ERK in the epididymis is essential for male fertility. Proc Natl Acad Sci U S A, 2014, 111(52):18643-18648.
16.	setia s, Nehru B, sanyal sN. Upregulation of MAPK/Erk and PI3K/Akt pathways in ulcerative colitis-associated colon cancer. Biomed Pharmacother, 2014, 68(8):1023-1029.
17.	Russnes HG, Caldas C. eEF2K-a new target in breast cancers with combined inactivation of p53 and PTEN. EMBO Mol Med, 2014, 6(12):1512-1514.
18.	Neshat Ms, Mellinghoff IK, Tran C, et al. Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci U S A, 2001, 98(18):10314-10319.
19.	Jin CH, Wang AH, Chen JM, et al. Relationship between expression and prognostic ability of PTEN, sTAT3 and VEGF-C in colorectal cancer. Exp Ther Med, 2012, 4(4):633-639.
20.	litlekalsoy J, Hostmark JG, Costea DE, et al. Time-dependent biological differences in molecular markers of high-grade urothelial cancer over 7 decades (ras proteins, pTEN, uPAR, PAI-1 and MMP-9). Virchows Arch, 2012, 461(5):541-551.
21.	Chen Js, Wang Q, Fu XH, et al. Involvement of PI3K/PTEN/AKT/mTOR pathway in invasion and metastasis in hepatocellular carcinoma:Association with MMP-9. Hepatol Res, 2009, 39(2):177-186.
22.	stankovic T, Milinkovic V, Bankovic J, et al. Comparative analyses of individual and multiple alterations of p53, PTEN and p16 in nonsmall cell lung carcinoma, glioma and breast carcinoma samples. Biomed Pharmacother, 2014, 68(5):521-526.
23.	江志坚, 王小平. PTEN及nm23-H1蛋白表达在非小细胞肺癌转移中的意义.中国肺癌杂志, 2008, 11(6):793-797.
24.	Gauffin F, Diffner E, Gustafsson B, et al. Expression of PTEN and sHP1, investigated from tissue microarrays in pediatric acute lymphoblastic, leukemia. Pediatr Hematol Oncol, 2009, 26(1):48-56.
25.	Petrocca F, lieberman J. Promise and challenge of RNA interferencebased therapy for cancer. J Clin Oncol, 2011, 29(6):747-754.
26.	Wang s, Cheng Z, Yang X, et al. Effect of wild type PTEN gene on proliferation and invasion of multiple myeloma. Int J Hematol, 2010, 92(1):83-94.
27.	sun Y, Tian H, Wang l. Effects of PTEN on the proliferation and apoptosis of colorectal cancer cells via the phosphoinositol-3-kinase/Akt pathway. Oncol Rep, 2015, 33(4):1828-1836.
28.	Das R, Bhattacharya K, sarkar s, et al. Mahanine synergistically enhances cytotoxicity of 5-fluorouracil through ROs-mediated activation of PTEN and p53/p73 in colon carcinoma. Apoptosis, 2014, 19(1):149-164.
29.	Marx J. Molecular biology. Cancer's perpetual source? Science, 2007, 317(5841):1029-1031.

1. Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin, 2011, 61(2):69-90.
2. shaukat A, Mongin sJ, Geisser Ms, et al. long-Term mortality after screening for colorectal cancer. N Engl J Med, 2013, 369(12):1106-1114.
3. 汪晓东, 李立.结直肠癌外科的新理念与挑战.中国普外基础与临床杂志, 2014, 21(6):782-786.
4. siegel R, Desantis C, Jemal A. Colorectal cancer statistics, 2014. CA Cancer J Clin, 2014, 64(2):104-117.
5. stambolic V, suzuki A, De la Pompa Jl, et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell, 1998, 95(1):29-39.
6. Ma JC, sawai H, Matsuo Y, et al. Interleukin-1alpha enhances angiogenesis and is associated with liver metastatic potential in human gastric cancer cell lines. J Surg Res, 2008, 148(2):197-204.
7. Ma JC, sawai H, Matsuo Y, et al. IGF-1 mediates PTEN suppression and enhances cell invasion and proliferation via activation of the IGF-1/PI3K/Akt signaling pathway in pancreatic cancer cells. J Surg Res, 2010, 160(1):90-101.
8. 张永红, 于冬冬, 何乐亚, 等.结肠癌原发灶及肝转移灶中PTEN蛋白的表达研究.中国组织化学与细胞化学杂志, 2010, 19(1):85-87.
9. 艾源.结直肠癌患者是否存在PTEN基因突变?中国普外基础与临床杂志, 2013, 20(7):811.
10. Hollander MC, Blumenthal GM, Dennis PA. PTEN loss in the continuum of common cancers, rare syndromes and mouse models. Nat Rev Cancer, 2011, 11(4):289-301.
11. 刘丹, 徐建洪, 焦泽龙, 等. PTEN基因功能及其调控研究进展.解放军医药杂志, 2014, 26(4):111-116.
12. Ma JC, sawai H, Matsuo Y, et al. IGF-1 and PTEN regulate the proliferation and invasiveness of colon cancer cells through opposite effects on PI3K/Akt signalling. Arch Med Sci, 2009, 2:195-206.
13. Ma JC, sawai H, Ochi N, et al. PTEN regulates angiogenesis through PI3K/Akt/VEGF signaling pathway in human pancreatic cancer cells. Mol Cell Biochem, 2009, 331(1-2):161-171.
14. Hempel N, Melendez JA. Intracellular redox status controls membrane localization of pro-and anti-migratory signaling molecules. Redox Biol, 2014, 20(2):245-250.
15. Xu BF, Washington AM, Hinton BT. PTEN signaling through RAF1 proto-oncogene serine/threonine kinase (RAF1)/ERK in the epididymis is essential for male fertility. Proc Natl Acad Sci U S A, 2014, 111(52):18643-18648.
16. setia s, Nehru B, sanyal sN. Upregulation of MAPK/Erk and PI3K/Akt pathways in ulcerative colitis-associated colon cancer. Biomed Pharmacother, 2014, 68(8):1023-1029.
17. Russnes HG, Caldas C. eEF2K-a new target in breast cancers with combined inactivation of p53 and PTEN. EMBO Mol Med, 2014, 6(12):1512-1514.
18. Neshat Ms, Mellinghoff IK, Tran C, et al. Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci U S A, 2001, 98(18):10314-10319.
19. Jin CH, Wang AH, Chen JM, et al. Relationship between expression and prognostic ability of PTEN, sTAT3 and VEGF-C in colorectal cancer. Exp Ther Med, 2012, 4(4):633-639.
20. litlekalsoy J, Hostmark JG, Costea DE, et al. Time-dependent biological differences in molecular markers of high-grade urothelial cancer over 7 decades (ras proteins, pTEN, uPAR, PAI-1 and MMP-9). Virchows Arch, 2012, 461(5):541-551.
21. Chen Js, Wang Q, Fu XH, et al. Involvement of PI3K/PTEN/AKT/mTOR pathway in invasion and metastasis in hepatocellular carcinoma:Association with MMP-9. Hepatol Res, 2009, 39(2):177-186.
22. stankovic T, Milinkovic V, Bankovic J, et al. Comparative analyses of individual and multiple alterations of p53, PTEN and p16 in nonsmall cell lung carcinoma, glioma and breast carcinoma samples. Biomed Pharmacother, 2014, 68(5):521-526.
23. 江志坚, 王小平. PTEN及nm23-H1蛋白表达在非小细胞肺癌转移中的意义.中国肺癌杂志, 2008, 11(6):793-797.
24. Gauffin F, Diffner E, Gustafsson B, et al. Expression of PTEN and sHP1, investigated from tissue microarrays in pediatric acute lymphoblastic, leukemia. Pediatr Hematol Oncol, 2009, 26(1):48-56.
25. Petrocca F, lieberman J. Promise and challenge of RNA interferencebased therapy for cancer. J Clin Oncol, 2011, 29(6):747-754.
26. Wang s, Cheng Z, Yang X, et al. Effect of wild type PTEN gene on proliferation and invasion of multiple myeloma. Int J Hematol, 2010, 92(1):83-94.
27. sun Y, Tian H, Wang l. Effects of PTEN on the proliferation and apoptosis of colorectal cancer cells via the phosphoinositol-3-kinase/Akt pathway. Oncol Rep, 2015, 33(4):1828-1836.
28. Das R, Bhattacharya K, sarkar s, et al. Mahanine synergistically enhances cytotoxicity of 5-fluorouracil through ROs-mediated activation of PTEN and p53/p73 in colon carcinoma. Apoptosis, 2014, 19(1):149-164.
29. Marx J. Molecular biology. Cancer's perpetual source? Science, 2007, 317(5841):1029-1031.

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Effect of PTEN siRNA on Proliferation and Invasion in Colon Cancer Cells

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