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From the Division of Gastroenterology,*
and the
Department of Oncology,§
Johns Hopkins
University School of Medicine, Baltimore, Maryland; and the Institute
of General and Experimental Pathology,
and
Institute of Clinical Pathology,
the
University of Vienna, Vienna, Austria
Previous reports indicate that the mRNA for the cardiac isoform of
the voltage-gated L-type calcium channel (
1C) is
elevated in colon cancer. The aim of these experiments was to verify
that the mRNA for 1C was significantly increased in
tumors of two separate populations of patients when compared to normal
adjacent mucosa. The second aim was to measure the distribution of
1C using immunocytochemistry in normal human colon and
in colon cancer and to determine what might regulate the channel
expression. Biopsies were taken from patients with various stages of
colon cancer and nearby normal mucosa were used as control. RNA was
prepared and mRNA level measured by semiquantitative reverse
transcriptase-polymerase chain reaction. The mRNA of the calcium
channel was compared with other markers including ß-actin. The mRNA
for 1C was increased significantly in colon cancers
compared to nearby adjacent mucosa. Using confocal microscopy
1C was localized mainly at the apical membrane in the
surface epithelium of normal human colon with less distribution on the
lateral and basal membranes. The channel was localized on the lateral
and basal membranes in crypt cells. Calcium channel localization
appeared to be nearer nuclei in colon cancer samples, in part
because of the smaller size of the cells. Likewise, cultured
Caco-2 and T84 cells showed a membrane distribution. Western blotting
indicated that 1C protein was increased in nonconfluent
cultures of colonic carcinoma cells compared to confluent cells and
immunocytochemistry confirms that there is more calcium channel protein
in cells that are nonconfluent. We conclude that the increase in mRNA
of 1 subunit of the cardiac isoform of the L-type
calcium channel may be a useful marker of colon cancer compared to
other markers because the increase is large and this increase can be
documented on small samples using a simple semiquantitative reverse
transcriptase-polymerase chain reaction. We found that
1C protein is increased when colonic cells are
nonconfluent or dividing which may account for the increase in
cancer.
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