Expression of the fragile histidine triad gene in normal rat tissues and human kidney cancer cell lines.
The fragile histidine triad (FHIT) gene located in human chromosome 3p14.2 is a candidate tumor suppressor gene that has a diadenosine triphosphate (Ap3A) hydrolase activity, but its role in carcinogenesis remains uncertain. To investigate the role of FHIT in normal cells, specific polyclonal antibodies to recombinant rat FHIT protein were generated. Immunoblot analysis revealed the 17-kd FHIT protein was most abundantly expressed in kidney and liver, whereas heart, skeletal muscle, and adrenal gland expressed trace amounts. In kidney, immunohistochemical staining was strongly observed in distal convoluted tubule and collecting duct during postnatal growth period. By a nested reverse transcription-PCR analysis of FHIT from 2 human kidney cancer cell lines, four abnormal-sized FHIT transcripts, with deletion and/or insertion, were detected. These were derived from the results of exon skipping, and/or insertion of FHIT intron 5 sequence, or selection of cryptic splice site within the FHIT cDNA sequence 179-180. Taken together, our data indicate that FHIT expression is frequently altered in human kidney cancer cell lines by alternative splicing, and suggest that the FHIT protein may play a pivotal role in regulating intracellular metabolism of the distal convoluted tubule and collecting duct in maturity.
Lee
Sang-Han
SH
Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheon-An 330-090, Korea. L1037624@sch.ac.kr
Kim
Tae-Jung
TJ
Kim
Ho-Young
HY
Kim
Jin-Gook
JG
Park
Mi-Kyoung
MK
Woo
Kee-Min
KM
Kim
Sung-Ho
SH
Hwang
Kyu-Yoon
KY
Nam
Hae-Seon
HS
Min
Young-Ki
YK
Cho
Man-Hee
MH
D017766:Acid Anhydride Hydrolases / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002847:Chromatography, Agarose; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism; D005091:Exons / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D007680:Kidney Neoplasms / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099170:Bis(5'-adenosyl)triphosphatase
D017766:Acid Anhydride Hydrolases; D017931:DNA Primers; D018076:DNA, Complementary; D005978:Glutathione; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; D000099170:Bis(5'-adenosyl)triphosphatase