Estimation of S-phase fraction in tumor tissue sections by immunohistochemical staining of PCNA.
We describe a method for measuring the size of the S-phase fraction in human tumor tissue sections using an antibody to PCNA (PC10). Although treatment with Triton X-100 before fixation extracted a large amount of PCNA from the cells even in frozen tissue sections, PCNA remained exclusively in S-phase cells. Immunohistochemical staining of PCNA after the detergent treatment allowed estimation of the S-phase fraction in solid tumors. The validity of the method was directly proven by double staining of bromodeoxyuridine (BrdU) and PCNA in HeLa cells. The PCNA-positive cells were identical with BrdUrd-positive cells after the detergent treatment. In contrast, almost all HeLa cells in the exponentially growing phase were positive for PC10 without treatment with Triton X-100.
Sasaki
K
K
Department of Pathology, Iwate Medical University, Morioka-shi, Japan.
Kurose
A
A
Ishida
Y
Y
Matsuta
M
M
D005260:Female; D005833:Genital Neoplasms, Female / Q000473:pathology; D006367:HeLa Cells / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D009369:Neoplasms / Q000473:pathology*; D009687:Nuclear Proteins / Q000032:analysis*; D017830:Octoxynol / Q000494:pharmacology; D018809:Proliferating Cell Nuclear Antigen; D016196:S Phase
D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D017830:Octoxynol