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Robert Disselkamp
Solis Research
Richland, WA 99352 USA
thegreenphd@gmail.com
ABSTRACT
All cells exist in an equilibrium state between their intracellular and extracellular fluid
concentrations, related through osmotic pressure. The concentration of normal human
cells is approximately 0.31 osmolar. Furthermore, it has been known for 30 years that
cancerous cell tissue contains about 0.11 osmolar more non-inorganic (e.g., debris)
material than healthy human cells, thus are at about 0.42 osmolar. This difference is
proposed here to be utilized as a cancer therapy with specific treatment. This study is a
literature review and paper study. No clinical or experimental work was performed. The
premise here is that by dosing of a benign agent, this will lead to cancer cell extreme
cytosis followed by aptosis. This is because by increasing the intracellular concentration
within cancer cells compared to their extracellular concentration, even more than they
exist naturally, can cause cancer cell death. Hence, cancer cells may be made to swell and
rupture compared to normal tissue, in vivo, due to the osmolarity values always being
greater for cancer cells. A novel cancer therapy based on mannitol dosing is proposed
here. The mechanism proposed here is that extreme mannitol dosing (e.g., or other
suitable benign agent) will preferentially lead to cancer cell aptosis. An unsustained
intracellular osmotic pressure causing endocytosis (about 22 atmospheres), leading to
preferential cancer cell apoptosis, is proposed.
Keywords
Osmolysis; Lysis; Osmotic pressure; Extracellular and intracellular; Amino acids;
Mannitol
1. Introduction
The basic chemical composition of cancer, and normal cells, has been known for more
than 30 years. This is true in regard to the primary components, such as inorganic ions,
amino acids, vitamins, and glucose [1-2]. More recent to our understanding, however, is
the composition and function of the molecular biology (proteins, enzymes, transport
processes, etc.) of both normal and cancerous cell tissue. Phenomenal advancements in
cancer research have occurred, where largely the mechanism in chemotherapy is making
use of selective binding sites (proteins) whereby anti-tumor agents can be attached.
Most of the chemotherapy cancer treatment options are cancer type specific. There are
some universally applicable treatment methods, based upon a single mechanism of action
across the broad cancer types that exist. However, no cancer treatment options based on
extreme endocytosis currently exist. In this work, a general mechanism is discussed based
on a therapy involving cancer cell extreme endocytosis, resulting in preferential cancer
cell rupture by an increased osmotic pressure resulting from an added benign solute to the
treated normal and cancer cell human system (e.g., patient). As is discussed here, the
reason for preferential cancer cell endocytosis and necrosis, compared to normal cells, is
two-fold. First, cancer cells contain debri solutes in a greater concentration compared to
healthy cells. Second, by introducing a dosing therapy based on increasing intracellular
concentrations of both healthy and cancer cells, such as by using mannitol (or similar
benign solute material) this can preferentially rupture cancer cells due their inherent
greater osmotic concentration. The mechanism of cancer cell necrosis arises from more
intracellular debris and solute material compared to healthy cells plus the added benign
solute mannitol dosing. It could be that this approach could be used not only in cancer
treatment, but also as a cancer preventative measure.
The study of human and animal cell osmotic pressure research is not new. Pioneering
work by Eagle [1] led the way in our understanding how extracellular (amino acid)
concentrations effect cancer cell growth. Subsequent work by Steenbergen, Hill, and
Jennings [2], and Malek and co-workers [3] demonstrated how healthy cells undergo
endocytosis when exposed to mannitol solutions in an extracellular matrix. As of
relevance here, work ranging from hemolysis [4], cancer cell extracellular and
intracellular work [5], extreme dosing mannitol therapy in head trauma [6], a solute
toxicity study [7], and virology [8], all pertain to the relationship between extracellular
and intracellular equilibrium manifested in hypotonic, isotonic, or hypertonic systems.
An examination of much of the references of this work show our knowledge of cancer
and healthy cell composition and endocytosis is not new. However, in this manuscript the
concept of exploiting the increased cancer cell solute osmolarity to cause necrosis in a
new cancer treatment is novel and the purpose of this report.
2. Methods and Materials
This work is a theoretical study based on published scientific literature from the medical
field. This work is not based on human, animal, or clinical studies. The principle of
osmotic pressure is applied to the unique system of cancer cell biochemistry (Results
section) to estimate how cancer cell necrosis can occur extrapolated from prior published
work (Discussion section).
Discussion
Intracellular Cancer Cell Concentration and Inhibited Cellular Growth
A pioneering study by Eagle [1] was performed that quantified the growth rate of cancer
cells as a function of concentration of amino acids in the growth medium. His chosen
type of cancer cell tissue was the human carcinoma (strain HeLa) tissue cultures.
Although not initially obvious from his study, was a relationship showing his cellular
multiplication factor (e.g., quantifying cell growth) versus various specific amino acids at
their concentrations. Figure 1 here presents a reanalysis of his data, and this serves as a
foundation for this work.
Property
Steenbergen et
Malek et al. [3]
al. [2]
(Osmolar)
(Osmolar)
(Osmolar)
0.32 - a
0.60 - d
0.15 - d
(0.03)
(0.30 - d)
(0.00 - d)
Extracellular
Total
(mannitol
portion -b)
Intracellular
Apoptosis (%)
0%
42 % - e
4.7 %
a Contains contributions from Krebs-Ringer phosphate (0.29 osmolar) plus
mannitol (0.03 osmolar).
b Included in total extracellular concentration.
c Taken from Figure 1 (Steenbergen et al., ref. [2]).
d The control used was a phosphate-buffered saline (PBS) solution at 0.30
osmolar.
e When mannitol at 0.30 osmolar was replaced with NaCl or urea, at 0.30
osmolar, the apoptosis rates were
42% and 31%, respectively. Again the PBS control was used at 0.30
osmolar, for a total extracellular concentration of 0.60 osmolar.
It is important to note in the data of Table 2 that before mannitol dosing is such that the
extracellular and intracellular concentrations are in an isotonic state. Indeed, this
requirement is not new, as work by Eagle and Brewer dating to the 1920s has shown this
to be the case [4]. It is seen from Table 2 that the work of Steenbergen et al. [2] illustrates
an uptake in mannitol, preferentially enters the cell so that its intracellular is 0.42-0.32 =
0.10 osmolar larger than its extracellular concentration. Hence, the preferential uptake is
0.10 osmolar (uptake)/0.03 osmolar (mannitol exposure) = 3.3-fold increase. This is key
because it shows that mannitol is preferentially concentrated within the cell (e.g.,
intracellular). Although cell swelling was observed by Steenbergen et al. [2], no
endocytosis induced apoptosis occurred.
The final two columns of Table 2 presents work by Malek et al. [3], where a larger
exposure to mannitol at 0.30 osmolar (with total extracellular concentration) of 0.60
osmolar, yielded a 42% apoptosis rate caused by endocytosis. A somewhat severe
hypotonic environment of the cells presented as the final column of Table 2 (without
mannitol), where the extracellular concentration is 0.15 osmolar leads to apoptosis in only
4.7% of the examined cells. Hence the preferential uptake of mannitol is important only to
choose its use as a chemical solute to increase the patients total osmolarity. The
hypertonic state of the solution, and even more so the preferential uptake of mannitol by
the cells, is seen to be the driving force for extreme cell endocytosis and apoptosis. As
shown by Malek et al. [3], either NaCl or urea can also be effective in causing cell
apoptosis, however as a cancer therapy they would not be well tolerated in patients at
these large concentrations, and hence are not considered as treatment options.
Conclusions
In this paper the known larger osmolarity of cancer cells compared to healthy cells is
exploited in a proposed patient therapy whereby a patient is given a benign solute at high
concentration, such a mannitol. This causes an increase in both cancer and healthy cell
osmolarity, however because of the increased cancer cell original osmolarity in excess of
that of healthy cells, the osmotic pressure of the cancer cells will be larger and increase
with mannitol dosing causing extreme endocytosis. This is a novel cancer treatment
therapy not previously proposed, despite the fact it has been known for more than 30 years
that cancer cells contain a greater osmolarity than healthy cells. The efficacy of the use of
extreme mannitol dosing therapy to cause cancer cell endocytosis and rupture needs
further study to demonstrate its viability among the various types of cancers that treatment
options that exist.
References
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JMJ