Role of protein kinase Cµ isoform expression in dimethylhydrazine-induced vascular endothelial proliferation

Nam,Subong; Choi,Soojong; Lee,Jaewoo; Kim,Joohyoung; Song,Jisun; Bae,Yongchan

doi:10.3892/mmr.2012.932

Role of protein kinase Cµ isoform expression in dimethylhydrazine-induced vascular endothelial proliferation

Authors:
- Subong Nam
- Soojong Choi
- Jaewoo Lee
- Joohyoung Kim
- Jisun Song
- Yongchan Bae
View Affiliations

Affiliations: Department of Plastic and Reconstructive Surgery, School of Medicine, Pusan National University, Busan 602-739, Republic of Korea, Medical Research Institute, Pusan National University, Busan 602-739, Republic of Korea
Published online on: May 29, 2012 https://doi.org/10.3892/mmr.2012.932
Pages: 399-404

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

1,2-Dimethylthdrazine (DMH) has been known to induce vascular neoplasms, such as malignant endothelioma, in animal experiments through the induction of abnormal proliferation of human umbilical vein endothelial cells (HUVECs). We studied the effect of protein kinase C (PKC) isoforms on DMH-induced abnormal proliferation of vascular endothelium to identify the isoforms with higher relevance. The study was conducted with pure culture HUVECs in a control group and a 1x10-9 M DMH-treated group. The mRNA and protein expression of PKC isoforms in DMH-treated HUVECs was evaluated by reverse transcription-polymerase chain reaction and western blotting. DMH-induced PKC production was detected by a PKC activity assay. To investigate the role of the PKC isoforms in DMH-induced abnormal HUVEC proliferation, we modulated PKCµ expression in DMH-treated HUVECs using small interfering RNA. We determined the expression of 11 PKC isoforms by PCR analysis in both the control and DMH-treated groups, and the results were statistically analyzed to detect any differences. According to the results, both groups expressed 6 out of 11 isoforms. Expression of PKCµ was significant in the DMH-treated group, and downregulation of PKCµ inhibited DMH-induced abnormal HUVEC proliferation. The PKCµ isoform is believed to be important in the abnormal growth of vascular endothelial cells induced by DMH, and this was confirmed by an objective siRNA experiment, which showed a clear decrease in PKCµ expression. Therefore, it is believed that PKCµ is a key factor in the abnormal proliferation of vascular endothelial cells, and these results can be used as fundamental research data for abnormal vessel development or the embryologic mechanisms of vessel development.

View Figures

View References

1	Klagsbrun M and D’Amore PA: Regulators of angiogenesis. Annu Rev Physiol. 53:217–239. 1991. View Article : Google Scholar
2	Folkman J and Shing Y: Angiogenesis. J Biol Chem. 267:10931–11934. 1992.
3	Folkman J: What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst. 82:4–6. 1990. View Article : Google Scholar : PubMed/NCBI
4	Ingber D, Fujita T, Kishimoto S, Sudo K, Kanamaru T, Brem H and Folkman J: Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumour growth. Nature. 348:555–557. 1990. View Article : Google Scholar : PubMed/NCBI
5	Weidner N, Semple JP, Welch WR and Folkman J: Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med. 324:1–8. 1991. View Article : Google Scholar : PubMed/NCBI
6	Mulliken JB: Pathogenesis of hemangiomas. Vascular Birthmarks: Hemangiomas and Malformations. Mulliken JB and Young AE: Saunders; Philadelphia: pp. 63–76. 1988
7	Druckrey H: Production of colonic carcinomas by 1,2-dialkylhydrazines and azoalkanes. Carcinoma of the Colon and Antecedent Epithelium. Burdette WJ: Sprinpfield, III: Charles C Thomas; pp. 267–279. 1970
8	Druckrey H, Preussmann R, Matzkies F and Ivankovic S: Selective production of intestinal cancer in rats by 1,2-dimethylhydrazine. Naturwissenschaften. 54:285–286. 1967.PubMed/NCBI
9	Wiebecke B, Löhrs U, Gimmy J and Eder M: Production of tumors in the intestines of mice by 1,2-dimethylhydrazine. Z Gesamte Exp Med. 149:277–278. 1969.PubMed/NCBI
10	Kim HO, Kang YS, Bae YC, Park SY, Hwang SM and Nam SB: Gene expression profiling of 1,2-Dimethylhydrazine-stimulated human umbilical vein endothelial cells. J Korean Soc Plast Reconstr Surg. 31:858–864. 2004.
11	Kim SH, Kang YS, Bae YC, Park SY and Nam SB: RT-PCR of Up-Regulated Factors in Abnormally Proliferated Vascular Endothelial Cells by 1,2-Dimethylhydrazine. J Korean Soc Plast Reconstr Surg. 32:689–698. 2005.
12	Bae YC, Park SY, Nam SB, Herh JY and Kang YS: A study for the mechanism of abnormal proliferation in vascular endothelial cells using inhibitors to the signal transduction pathway. J Korean Soc Plast Reconstr Surg. 33:5–12. 2006.
13	Lee J, Bae YC, Park SY, Moon JS and Nam SB: Role of protein kinase C in abnormal proliferation of vascular endothelial cell induced by 1,2-dimethylhydrazine; analysis of isoform. J Korean Soc Plast Reconstr Surg. 34:8–12. 2007.(In Korean).
14	Livneh E and Fishman DD: Linking protein kinase C to cell-cycle control. Eur J Biochem. 248:1–9. 1997. View Article : Google Scholar : PubMed/NCBI
15	Nishi K, Schnier JB and Bradbury EM: The accumulation of cyclin-dependent kinase inhibitor p27kip1 is a primary response to staurosporine and independent of G1 cell cycle arrest. Exp Cell Res. 243:222–231. 1998. View Article : Google Scholar : PubMed/NCBI
16	Fukumoto S, Nishizawa Y, Hosoi M, Koyama H, Yamakawa K, Ohno S and Morii H: Protein kinase C delta inhibits the proliferation of vascular smooth muscle cells by suppressing G1 cyclin expression. J Biol Chem. 272:13816–13822. 1997. View Article : Google Scholar : PubMed/NCBI
17	Ways DK, Kukoly CA, DeVente J, Hooker JL, Bryant WO, Posekany KJ, Fletcher DJ, Cook PP and Parker PJ: MCF-7 breast cancer cells transfected with protein kinase C-alpha exhibit altered expression of other protein kinase C isoforms and display a more aggressive neoplastic phenotype. J Clin Invest. 95:1906–1915. 1995. View Article : Google Scholar
18	Dean N, McKay R, Miraglia L, Howard R, Cooper S, Giddings J, Nicklin P, Meister L, Ziel R, Geiger T, Muller M and Fabbro D: Inhibition of growth of human tumor cell lines in nude mice by an antisense of oligonucleotide inhibitor of protein kinase C-alpha expression. Cancer Res. 56:3499–3507. 1996.PubMed/NCBI
19	O’Brian C, Vogel VG, Singletary SE and Ward NE: Elevated protein kinase C expression in human breast tumor biopsies relative to normal breast tissue. Cancer Res. 49:3215–3217. 1989.PubMed/NCBI
20	Schwartz GK, Redwood SM, Ohnuma T, Holland JF, Droller MJ and Liu BC: Inhibition of invasion of invasive human bladder carcinoma cells by protein kinase C inhibitor staurosporine. J Natl Cancer Inst. 82:1753–1756. 1990. View Article : Google Scholar : PubMed/NCBI
21	Newton AC: Protein kinase C: structure, function, and regulation. J Biol Chem. 270:28495–28498. 1995. View Article : Google Scholar : PubMed/NCBI
22	Gampper TJ and Morgan RF: Vascular anomaly: Hemangiomas. Plast Reconstr Surg. 110:572–585. 2002. View Article : Google Scholar
23	Mulliken JB and Glowacki J: Hemangiomas and vascular malformations in infants and children: A classification based on endothelial characteristics. Plast Reconstr Surg. 69:412–422. 1982. View Article : Google Scholar : PubMed/NCBI
24	Clemens MJ, Trayner I and Menaya J: The role of protein kinase C isoenzymes in regulation of cell proliferation and differentiation. J Cell Sci. 103:881–887. 1992.PubMed/NCBI
25	Bischoff J: Perspectives series; cell adhesion in vascular biology. J Clin Invest. 99:373–376. 1997.
26	Lee OH, Kim YM, Lee YM, Moon EJ, Lee DJ, Kim JH, Kim KW and Kwon YG: Sphingosine1-phosphat induces angiogenesis: its angiogenic action and signaling mechanism in human umbilical vein endothelial cells. Biochem Biophys Res Commun. 264:743–750. 1999. View Article : Google Scholar : PubMed/NCBI
27	Lutz MP, Esser IB, Flossmann-Kast BB, Vogelmann R, Luhrs H, Friess H, Buchler MW and Adler G: Overexpression and activation of the tyrosine kinase Src in human pancreatic carcinoma. Biochem Biophys Res Commun. 243:503–508. 1998. View Article : Google Scholar : PubMed/NCBI
28	Hug H: Protein kinase C isoenzymes divergence in signal transduction? Biochem J. 291:329–343. 1993.PubMed/NCBI
29	Yang JH and Kodavinti PR: Possible molecular targets of halogenated aromatic hydrobons in neural cells. Biochem Biophys Res Commun. 280:1372–1377. 2001. View Article : Google Scholar : PubMed/NCBI
30	Mischak H, Goodnight JA, Kolch W, Martiny-Baron G, Schaechtle C, Kazanietz MG, Blumberg PM, Pierce JH and Mushinski JF: Overexpression of prtein kinase C-δ and -ɛ in NIH 3T3 cells induces opposite effects on growth, morphology, anchorage dependence, and tumorigeniccity. J Biol Chem. 268:6090–6096. 1993.
31	Grabam MA, Rawe I, Dartt DA and Joyce NC: Protein kinase C refulation of corneal endothelial cell proliferation and cell cycle. Invest Ophthalmol Vis Sci. 41:4124–4132. 2000.PubMed/NCBI
32	Farivar RS, Gardner-Thorpe J, Ito H, Arshad H, Zinner MJ, Ashley SW and Whang EE: The efficacy of tyrosine kinase inhibitors on human pancreatic cancer cell lines. J Surg Res. 115:219–225. 2003. View Article : Google Scholar : PubMed/NCBI