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.	汪曉東, 李立.結直腸癌外科的新理念與挑戰.中國普外基礎與臨床雜志, 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.	張永紅, 于冬冬, 何樂亞, 等.結腸癌原發灶及肝轉移灶中PTEN蛋白的表達研究.中國組織化學與細胞化學雜志, 2010, 19(1):85-87.
9.	艾源.結直腸癌患者是否存在PTEN基因突變?中國普外基礎與臨床雜志, 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.	劉丹, 徐建洪, 焦澤龍, 等. PTEN基因功能及其調控研究進展.解放軍醫藥雜志, 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.	江志堅, 王小平. PTEN及nm23-H1蛋白表達在非小細胞肺癌轉移中的意義.中國肺癌雜志, 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. 汪曉東, 李立.結直腸癌外科的新理念與挑戰.中國普外基礎與臨床雜志, 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. 張永紅, 于冬冬, 何樂亞, 等.結腸癌原發灶及肝轉移灶中PTEN蛋白的表達研究.中國組織化學與細胞化學雜志, 2010, 19(1):85-87.
9. 艾源.結直腸癌患者是否存在PTEN基因突變?中國普外基礎與臨床雜志, 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. 劉丹, 徐建洪, 焦澤龍, 等. PTEN基因功能及其調控研究進展.解放軍醫藥雜志, 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. 江志堅, 王小平. PTEN及nm23-H1蛋白表達在非小細胞肺癌轉移中的意義.中國肺癌雜志, 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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