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Differences in the Intracellular Concentration of Elements in Normal and Cancerous Liver Cells as Determined by X-ray Microanalysis

Department of Anatomy, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78284

Elemental X-ray microanalysis was performed on hepatocytes of normal, young adult A/J mice and on host hepatocytes and tumor cells of mice that had been given injections in the left flank of transplantable H6 hepatoma cells. After the mice were killed by cervical dislocation, tissue was rapidly removed, mounted on tubular brass holders with minced liver, and frozen in liquid propane. The frozen tissue was sectioned at 4 µm in a cryostat, freeze-dried, and electron probed in a scanning electron microscope. X-ray spectra were collected and processed with an energy-dispersive detector and pulse-height analysis system. Data were collected from ten cells (nucleus and cytoplasm) from each of six data sets (two animals each for normal hepatocytes, host hepatocytes, and hepatoma cells). Six elements of biological significance were detected: sodium, magnesium, phosphorus, sulfur, chlorine, and potassium. Normal hepatocytes and host hepatocytes differed very little in their elemental contents; the latter did show a decrease in magnesium from normal hepatocytes. However, the sodium and chlorine concentrations of hepatoma cells were more than double those of normal hepatocytes. These findings are consistent with the hypothesis that a high sodium concentration associated with a low transmembrane potential is mitogenic. The only elements showing an unequal distribution between nucleus and cytoplasm were chlorine and sulfur. Chlorine had a nuclear/cytoplasmic ratio of significantly less than unity in normal and host hepatocytes and greater than unity in transformed hepatoma cells. For sulfur a slight but significant concentration in the nucleus versus the cytoplasm occurs in normal and host hepatocytes but not in H6 hepatoma cells.

¹ Recipient of NIH Institutional Grant SO1 RR 05654.

² To whom requests for reprints should be addressed.

³ Recipient of USPHS Grant CA16831 from the National Cancer Institute.

Received 12/12/77. Accepted 3/28/78.

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