SPECIFIC INHIBITION BY L-GLUCOSE OF CANCER CELL PROLIFERATION POTENTIATION OF HYPERTHERMIA, RADIATION, AND CHEMOTHERAPY EFFECTS. ~ Haim I. Bicher, M.D.

~, Samuel Yarmenko, Ph.D.+, Adolf Weinson, M.D.+, Irene Surowiec, Ph.D.~, and Nodar Mitagvaria, Ph.D.#. Valley Cancer Institute, Los Angeles, California, USA Moscow Cancer Research Center, Russian Federation Physielogy Institute, Tbilisi, Georgia Cancer cells are known to consume large amounts of glucose as a source of energy permitting the exaggerated use of amlno acids and nucleosides in the synthesis of DNA. Further, it has been shown that many cancers are almost entirely de~endant on glucose for their energy requirements(l). Exploiting this trait by supplying excess glucose, the aerobic glycolysis of tumor cell can be stimulated in vivo to a very high extent. However due to substrate deficiencies, it appears that many cancer cells take in much more glucose thanthey can metabolize efficiently and, as a consequence, excrete large amounts of lactic acid. This leads to a lower pH value in the malignant cells which can increase the susceptibility of the tumor to various therapies (2). In prior prelimlnary publications (3), we have described the surprising fact that the L isomer of Glucose ( L - Glucose) inhibits cancer cell growth Therefore we undertook to evaluate the effect of L. glucose on neoplastic as wel! as healthy cells. Furthermore experiments were conducted to elucidate the possible potential enhancement to other treatment modalities( Hyperthermia, Radiation and Chemotherapy) by the use of this substance.
MATERIALS AND METHODS " 1. Cell lines - Experiments were performed on normal human embryo intestinal cells grown on 199 - !0 % calf serum media, gentamicl~n treated. Cells are grown under 02 95% - CO2 10% for 14 days. L or D Glucose added for 48 hours. Colonies were compared after 14 days. Similar experiments were performed on the following tumor cell lines: Colon Adenocarc!noma grown in RPMI 1640 media. Rat mammary tumor cells (gL-Glioma) in 199 - Fetal Calf media, Human ovarian cancer ce!ls in the same media, as well as HeLla S3 and human melanoma cells under the same conditions.

2. Drugs and Cytotoxic Methods - Hyperthermia for one hour at 43 C was achieved by placing the treated flasks in a regulated water bath, under oxic (90% 02) or anoxic (5% 02 - 95% CO2) conditions. L Glucose or D Glucose, 5 Fluoruracil (5FU) and Cis. Platinum were added in concentrations as indicated in results. Gamma irradiation was delivered in doses of 2 G in one hour using a 200 KV X Ray machine.

RESULTS: i. L-Glucose exhibits a cytostatic and cytocidal effect on cancer cells, but not on normal cells, as shown in Figure i. L-GLUCOSE DOES NOT INHIBIT GROWTH NORMAL CHO CELLS L-Glucose inhibits~rowth of 9-L Glioma tumor cells Figure ! - L-Glucose inhibits the growth of malignant cells (right) but non of normal cells. These experiments were done in media with or without normal concentrations of D-Glucose. 2. L-Glucose potentiates the cytotoxic and cytocidal effect of hyperthermla - specifically under hypoxic conditions.

Figure 2 Effect of L-Glucose on growth rate of Ca ovarian cells under oxic (left) and Hypoxic conditions. L-Glucose potentiates the cytotoxic effect of hyperthermia, twice as much under anoxic than oxic conditions.
3. L-Glucose potentiates the effectiveness of Gamma Irradiatlon.

Figure 3 - Effect of L-Glucose on potentiation inhibition of CaO cell growth by Gamma Irradiation. 4. L-Glucose potentiates the cytostatic effect of chemotherapeutic agents.

Figure 4 - Effect of L Glucose in potentiating the cytostatic effect of 5 FU on CA Ovarian cel! growth in Vitro.

The above experiments clearly demonstrate the potential therapeutic value o~ L- Glucose as an antineoplastic composition when used alone or in combination with other non-surgical cancer treatments. Further experimentation in vivo, as well as through toxicity experiments will of course be needed prior to human tests. The mode by which L-Glucose influences cancer cell survival is unclear. Obviously normal cells seem to be able to utilize or degrade the substance without impediment, while neoplastic cells are not (4). Also, neoplastic cells seem to be unable to utilize L-Glucose "as a source of energy ~o satisfy their increased metabolic needs. Probably, the potential use of the drug wi!! be more as an adjunct to chemo or radiation although the mechan!sm of action of this potentiation is yet to be determined and also unclear.

REFERENCES 1. Streffer, C.; Steinberg, F. "Effects on Glucose and Oxygen Metalbolism as We!! as Cell Proliferation after Treatment of Tumors with X-Rays, Heat and Sensitizer". Proc, 6th Intnl. Cng. on Hy~athermic Oncology vol. ! Dietzel-F, Basic Princioles in Hyoerthermic Tumor Therapy, Recent-Results-Cancer-Res,vol. 86, iss.8 (1983>: 177-90. Gi!lette-E-L, "Clinical Use Combined of Thermal Enhancement ~ ~w~. Therapeutic Gain for Hyperthermla with Radiation and or Drugs" 48418. Cancer -Res. volo44, iss.10 Suppl (1984 Oct>: 48368-

/. Dwarkanath, B.S. Enger~ Linked Modifications of the Radiation Response in a Human Cerevral Glioma Cell Line". 17 May 1989.
~ Vo!k, T.; "Effect of Glucose-Mediated pH Reduction and Cyclophoshamide on Oxygenation of Transplanted Rat Tumors, 31 Jul. 1992,

, Kim, J.L.; H]pox~c Ce!l Radiosensitization by Moderate Hyperthermia amd Glucose Deprivation"; Radiation Research93, 416~20 (1983). / Bicher, H.; Yarmonenko, S.; Wainson, S.; Surowiec, I,;-~Metagvaria, N. "Specific Inhibition of L-Glucose Cell Proliferation. Potentiation of Hyperthermia, Radiation, and Chemotherapy Effects"(Abstract). Proceeding of the i6th ISCH. Kyoto, Japan, June 13-16, 1993.