Establishment of a new OSCC cell line derived from OLK and identification of malignant transformation-related proteins by differential proteomics approach
Authors: Yanguang Zhang
Xu Guo
Liyan Xue
Weimin Zhang
Zhuwen Chen
Qimin Zhan
Liyuan Yu
Fei Fang
Jia Li
Caiyun Liu
Xiandong Yang
Jing Xiao
Xiaoyan Ma
Ming Fu
Lijia Dong
Lihua Zhang
Qun Zhao
Xuefeng Liu
Shujuan Shao
Guowu Ma
Yan Dong
Journal: Scientific Reports (2015)
Discipline: Multidisciplinary
Keywords: Oral squamous cell carcinoma; oral leukoplakia; carcinogens; B(a)P; DMBA
Affiliations: 1] College of Stomatology, Dalian Medical University, Dalian 116044, China [2] State Key Laboratory of Molecular Oncology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China., Dalian Institute of Chemical Physics, Chinese Academy of Science, Dalian 116023, China., College of Stomatology, Dalian Medical University, Dalian 116044, China., Institute of Cancer Stem Cell, Second Affiliated Hospital, Cancer Center, Dalian Medical University, Dalian 116044, China., State Key Laboratory of Molecular Oncology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China., Department of Engineering Mechanics, Dalian University of Technology, Dalian 116023, China., Department of Pathology, Cancer Hospital and Cancer Institute, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.

Abstract:  Oral squamous cell carcinoma (OSCC) is usually preceded by the oral premalignant lesions, mainly oral leukoplakia (OLK) after repeated insults of carcinogens, tobacco. B(a)P and DMBA are key carcinogens in tobacco smoke. In the present study, for the first time we established the cancerous cell line OSCC-BD induced by B(a)P/DMBA mixture and transformed from dysplastic oral leukoplakia cell line DOK. Cell morphology, proliferation ability, migration ability, colony formation, and tumorigenicity were studied and confirmed the malignant characteristics of OSCC-BD cells. We further identified the differential proteins between DOK and OSCC-BD cells by stable isotope dimethyl labeling based quantitative proteomic method, which showed 18 proteins up-regulated and 16 proteins down-regulated with RSD &lt; 8%. Differential proteins are mainly related to cell cycle, cell proliferation, DNA replication, RNA splicing and apoptosis. Abberant binding function, catalysis activity and transportor activity of differential proteins might contribute to the malignant transformation of OLK. Of the 34 identified differential proteins with RSD &lt; 8%, 13 novel cancer-related proteins were reported in the present study. This study might provide a new insight into the mechanism of OLK malignant transformation and the potent biomarkers for early diagnosis, meanwhile further facilitate the application of the quantification proteomics to carcinogenesis research. 

Retracted: No

PubPeer comments:
 Figure 5 of this paper shows two mice, one of which carries a large tumor on it side, created by injecting an oral cancer cell line. The text states that the tumors were about 1 cm in size - but they appear to be much larger on the photo. Could the authors clarify how big (mean and range) the tumors were obtained for this study? Were these in line with animal ethics guidelines? ![file] , 
 I am no expert on animal ethics, but I would certainly appreciate the authors sharing the ethics approval documentation as well as a copy of the u201cguidelinesu201d and u201cprotocolsu201d referred to in the text. Chinese language documents will suffice. u201c The animal experiments were carried out in accordance with the approved guidelines and all experimental protocols were approved by the Animal Care and Use Committee of Cancer Hospital and Cancer Institute in Chinese Academy of Medical Sciences and Peking Union Medical College.u201d , 
 #1 By Jove! Dr. Bik, itu2019s a whopper! While it is not something I particularly want to gloat about, one of mine from this particular publishing outfit is a soupu00e7on more necrotic. https: pubpeer.com publications A7E4AE815775161947341A3C63FC52 Given that Springer-Nature now have a research ethics office, which they didnu2019t see the need for in 2015, I recommend contacting the office directly. They have promised to do better in future. Contrast with PLOS One, which has retracted the two unethical xenografts that I anonymously reported to them: Bad Moon Rising https: pubpeer.com publications 6FCE06A14C3AC48194A0FFCEF0ACCB And https: pubpeer.com publications 40B3BF202DC5C813EAB8F4AAEC4BBA These ethical violations have nowhere to hide as weu2019ve moved to open access publication so that the whole world can see them and be horrified. Those who aspire to be scientists, wherever they are based, need to understand that ethical violations will now always backfire and that they are risking their careers. Keep up the good work. , 
 #1 Dear Dr. Bik, Thank you very much for your comments. The issue you brought up is very important. In this study, we developed an oral squamous cell carcinoma cell line OSCC-BD derived from an oral epithelial dysplasia cell line DOK and sought to evaluate the capacity of xenograft formation in vivo of these two cell lines. After a month of transplantation, we found that DOK cells did not form xenograft in nude mice, and OSCC-BD cells showed the heterogeneous xenograft formation ability in different nude mice. In the end of the experiments, we sacrificed the animals, excised xenografts and removed the surrounding soft tissues, and then measured the diameter of the isolated tumors by vernier caliper. The maximum diameters of all tumors did not exceed 2.0 cm, which was consistent with the Institutional Animal Care and Use Committee (IACUC) Guideline of many universities, including UCSF , University of Pennsylvania , Purdue University , et al. We would also like to say that during the animal experiments, we paid great attention to the animal welfare. We kept several lines in mind: 1. Experimental endpoint: The purpose of our experiment was to detect the tumor formation ability of DOK and OSCC-BD cells. To exclude the possibility of spontaneous tumorigenesis of DOK cells, we employed a month as the experimental period. 2. Humane endpoint: That is, in the process of the experiment, the overall wellu2010being of the animal took priority over the purpose of experiments, when we knew the experimental results, we timely selected the earlier stage as the end point of the experiment. 3. We set up the endpoint before the collapse of xenografts, which might cause discomfort in mice. 4. Tumor excision was performed after euthanasia, and all efforts were made to minimize suffering of animals. Once again, we highly appreciate your comments. , 
 #4 I appreciate the authoru2019s reply, of many words. Unfortunately, I am not seeing that the authors have responded to the specific concerns expressed in #1; for example; u201cCould the authors clarify how big (mean and range) the tumors were obtained for this study?u201d ,