Protein Expression Changes Induced by Cisplatin in an Oral Cancer Cell Line as Determined by Immunoprecipitation-Based High Performance Liquid Chromatography

다국어 초록 (Multilingual Abstract)

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

참고문헌 (Reference)

1 Fleitas T, "VEGF and TSP1levels correlate with prognosis in advanced non-small cell lung cancer" 15 : 897-902, 2013

2 McCormack ES, "Transforming growth factor-beta3 protection of epithelial cells from cycle-selective chemotherapy in vitro" 53 : 1149-1159, 1997

3 Liu P, "Transforming growth factor-beta and response to anticancer therapies in human liver and gastric tumors in vitro and in vivo" 16 : 599-610, 2000

4 Brauckmann C, "The interaction of platinum-based drugs with native biologically relevant proteins" 405 : 1855-1864, 2013

5 Li X, "Thalidomide down-regulates the expression of VEGF and bFGF in cisplatin-resistant human lung carcinoma cells" 23 : 2481-2487, 2003

6 Gasparini G, "Predictive and prognostic markers in a series of patients with head and neck squamous cell invasive carcinoma treated with concurrent chemoradiation therapy" 1 : 1375-1383, 1995

7 Ahmad S, "Platinum-DNA interactions and subsequent cellular processes controlling sensitivity to anticancer platinum complexes" 7 : 543-566, 2010

8 Calandrini V, "Platination of the copper transporter ATP7A involved in anticancer drug resistance" 43 : 12085-12094, 2014

9 Bozec A, "Molecular targeted therapies in the management of head and neck squamous cell carcinoma: recent developments and perspectives" 13 : 389-402, 2013

10 D’Amico TA, "Molecular biology of esophageal cancer" 10 : 451-469, 2000

11 Kartalou M, "Mechanisms of resistance to cisplatin" 478 : 23-43, 2001

12 Eljack ND, "Mechanisms of cell uptake and toxicity of the anticancer drug cisplatin" 6 : 2126-2133, 2014

13 Fleer EA, "Induction of resistance to hexadecylphosphocholine in the highly sensitive human epidermoid tumour cell line KB" 32A : 506-511, 1996

14 김연숙, "Immunoprecipitation-Based High Performance Liquid Chromatography Analysis of Human Mixed Saliva" 대한구강악안면병리학회 33 (33): 337-346, 2009

15 김연숙, "High Performance Liquid Chromatography Analysis of Human Salivary Protein Complexes" 대한구강악안면병리학회 38 (38): 381-388, 2014

16 Florescu-Zorila S, "H-7and fetal calf serum (FCS) act synergistically to increase apoptosis in the KB line of human oral carcinoma cells" 33 : 368-375, 2001

17 Yadav V, "Gatifloxacin induces S and G2-phase cell cycle arrest in pancreatic cancer cells via p21/p27/p53" 7 : e47796-, 2012

18 Wang YD, "Farnesoid X receptor antagonizes nuclear factor kappaB in hepatic inflammatory response" 48 : 1632-1643, 2008

19 Uno M, "Expression of cell cycle regulator genes in KB, a human squamous cell carcinoma cell line, after irradiation" 17 : 947-954, 2000

20 Stoika RS, "Effect of anticancer drugs on production of transforming growth factor and expression of p53 AND Bcl-2proteins by MCF-7 and T47D cell lines of human breast carcinoma" 30 : 35-41, 2008

21 Pal S, "Cytokines from the tumor microenvironment modulate sirtinol cytotoxicity in A549 lung carcinoma cells" 64 : 196-207, 2013

22 Herraez E, "Cisplatin-induced chemoresistance in colon cancer cells involves FXR-dependent and FXR-independent up-regulation of ABC proteins" 9 : 2565-2576, 2012

23 de Biasi AR, "Cisplatin-induced antitumor immunomodulation: a review of preclinical and clinical evidence" 20 : 5384-5391, 2014

24 Stachurska A, "Cisplatin up-regulates the in vivo biosynthesis and degradation of renal polyamines and c-Myc expression" 1689 : 259-266, 2004

25 Shen DW, "Cisplatin resistance: a cellular self-defense mechanism resulting from multiple epigenetic and genetic changes" 64 : 706-721, 2012

26 Dempke W, "Cisplatin resistance and oncogenes--a review" 11 : 225-236, 2000

27 Sedletska Y, "Cisplatin is a DNA-damaging antitumour compound triggering multifactorial biochemical responses in cancer cells:importance of apoptotic pathways" 5 : 251-265, 2005

28 Azuma M, "Cisplatin induces apoptosis in oral squamous carcinoma cells by the mitochondria-mediated but not the NF-kappaBsuppressed pathway" 39 : 282-289, 2003

29 Florea AM, "Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects" 3 : 1351-1371, 2011

30 이병주, "Change of the Expression of Human Telomerase Reverse TranscriptasemRNA and Human Telomerase RNA after Cisplatin and 5-FluorouracilExposure in Head and Neck Squamous Cell Carcinoma Cell Lines" 대한의학회 22 (22): 73-78, 2007

31 Harding J, "Cetuximab: an epidermal growth factor receptor chemeric human-murine monoclonal antibody" 41 : 107-127, 2005

32 Basu A, "Cellular responses to Cisplatin-induced DNA damage" 2010 : 201367-, 2010

33 Kondo N, "Antitumor effects of telomelysin in combination with paclitaxel or cisplatin on head and neck squamous cell carcinoma" 23 : 355-363, 2010

34 Pang R, "A study on effects of cisplatin and its mechanisms on human lung adenocarcinoma SLC-89 cells" 6 : 469-472, 2003

35 McCormack ES, "A p53growth arrest protects fibroblasts from anticancer agents" 2 : a37-a45, 1997

36 Crowe DL, "A common pathway for chemotherapy-induced apoptosis in human squamous cell carcinoma lines distinct from that of receptor mediated cell death" 23 : 2321-2328, 2003