title: Common housekeeping proteins are upregulated in colorectal adenocarcinoma and hepatocellular carcinoma, making the total protein a better "housekeeper".
abstract: Housekeeping proteins are essential endogenous controls for normalization as they are expected to be stably expressed. However, the stability of the expression level of housekeeping proteins needs to be assessed considering various experimental conditions. Our study evaluated the degree of variability of 7 commonly used housekeeping proteins with regard to their potential utility as normalizers in 56 pairs of matched colorectal adenocarcinoma (CRC) tissue samples and 6 pairs of hepatocellular carcinoma (HCC) tissue samples using multiple reaction monitoring (MRM) and Western blot analyses. A comprehensive experimental design and strict statistical analysis revealed that the expression levels of these 7 housekeeping proteins were not as stable as expected and they all exhibited upregulations to varying degrees in both the CRC and the HCC tissue samples. Consequently, we verified that using the amount of total protein instead of that of an individual protein can serve as a preferable control for studies of protein expression that require normalization.
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Hu
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Shujiao
authors.initials: S
authors.lastname: Du
authors.affiliation: Cancer Institute, Key Laboratory of Cancer Prevention and Intervention, The Second Affiliated Hospital, Zhejiang University, College of Medicine, Hangzhou, Zhejiang, 310009, PR China.
authors.forename: Jiekai
authors.initials: J
authors.lastname: Yu
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Xuhan
authors.initials: X
authors.lastname: Yang
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Chao
authors.initials: C
authors.lastname: Yang
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Daizhan
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Shengying
authors.initials: S
authors.lastname: Qin
authors.affiliation: School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, PR China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Yan
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Lin
authors.initials: L
authors.lastname: He
authors.affiliation: Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, 200240, PR China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Han
authors.affiliation: Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China.
authors.forename: Chunling
authors.initials: C
authors.lastname: Wan
mesh_terms: D000199:Actins / Q000378:metabolism; D000230:Adenocarcinoma / Q000175:diagnosis / Q000378:metabolism*; D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D004268:DNA-Binding Proteins; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism; D050810:Octamer Transcription Factors / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D016601:RNA-Binding Proteins / Q000378:metabolism; D012015:Reference Standards; D012269:Ribosomal Proteins / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D000199:Actins; D004268:DNA-Binding Proteins; C000609943:H3C2 protein, human; D006657:Histones; C083214:NONO protein, human; D034521:Nuclear Matrix-Associated Proteins; D050810:Octamer Transcription Factors; D016601:RNA-Binding Proteins; C489092:RPL19 protein, human; D012269:Ribosomal Proteins; C578986:TUBB1 protein, human; D014404:Tubulin; C041184:ribosomal protein S18; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: endogenous control; housekeeping protein; normalization; total protein amount; tumor