Re: Lung Cancer Treatment

[Guest guest]

" moonbeam " <moonbeam@...>

Subject: Lung Cancer Treatment

Date sent: Thu, 16 Aug 2001 06:41:02 -0600

> >>These findings lead us to believe that ozone--alone, in

> combination with radiation therapy (16), or in chemotherapy

> utilizing electrophilic compounds (17)--may have therapeutic

> value for patients with certain forms of lung cancer.

> Do you know which forms of lung cancer ozone helped?

Hi,

Here is the original article. Notice that lung, breast, uterine and

endometrial

cancers were inhibited by an active oxygen called medical Ozone (see references,

8,9,10

below). In my understanding, at least 98% of all tumor types are killed by

ozone. Its not

like a patented drug that only kills one type of cancer. Similarly at least 98%

of

viruses are killed by ozone. The reason is that viruses and cancers have no anti

oxidant

coating to protect them as good, aerobic cells do.

Ozone Selectively Inhibits Growth of Human Cancer Cells

Science Vol. 209, 22 Aug 1980, pp. 931-933

Abstract:

The growth of human cancer cells from lung, breast, and uterine

tumors was selectively inhibited in a dose-dependent manner by

ozone at 0.3 to 0.8 part per million of ozone in ambient air

during 8 days of culture. Human lung diploid fibroblasts served

as noncancerous control cells. The presence of ozone at 0.3 to

0.5 part per million inhibited cancer cell growth 40 and 60

percent, respectively. The non-cancerous lung cells were

unaffected at these levels. Exposure to ozone at 0.8 part per

million inhibited cancer cell growth more than 90 percent and

control cell growth less than 50 percent. Evidently, the

mechanisms for defense against ozone damage are impaired in

human cancer cells.

The effects of ozone on human health have been a focus of public

concern and scientific investigation for more than two decades

(I-4). Considerable attention has been devoted to assessing its

cellular effects (5) because it is the major constituent of the

ground-level oxidants in polluted air. Much has been learned

about the effects of ozone on normal tissue, but little is known

about its action on cancer cells. We have conducted experiments

in which continuous exposure to ozone at 0.3 ppm (6) selectively

inhibited the growth of human cancer cells 40 percent in 8 days.

Controlled levels of ozone (0.3 to 0.8 ppm) were continuously

generated by ultraviolet irradiation of filtered ambient air.

The ozonated air, containing 5 percent carbon dioxide, was

introduced at a constant flow rate of 4.0 liter/min into an

environmental chamber in an incubator maintained at 37 degrees

Celsius (Fig. 1). The ozone levels were assayed daily with a

spectrophotometric ozone analyzer. For comparison, noncancerous

human lung diploid fibroblasts (7) were cultured in the chamber

along with the cancer cells. The cancer cells were from

alveolar (lung) adenocarcinomas (8), breast adenocarcinomas (9),

uterine carcinosarcomas, and endometrial carcinomas (10). All

the cells were grown in 60-mm petri dishes in 10 ml of medium

and were placed in the chamber at the same time. Control cells

were incubated in an adjoining compartment receiving filtered

ambient air containing 5 percent carbon dioxide (4.0 liter/min).

Three petri dishes for each cell type were removed from each of

the two compartments every 48 hours for 8 days, and the number

of cells per plate were counted. All of the cancer cells showed

marked dose-dependent growth inhibition in ozone at 0.3 and 0.5

ppm (Fig 2.). There was no growth inhibition of the

noncancerous lung cells at these ozone levels, and they were

morphologically identical to the corresponding control cells.

At 0.8 ppm, the growth of the noncancerous cells was inhibited

50 percent, but all four types of cancer cells were inhibited

more than 90 percent.

After being cultured through 14 passages, the noncancerous cells

exhibited measurable growth inhibition and morphological changes

(vacuolation) in ozone at 0.5 ppm, suggesting that aging

increases the sensitivity of normal lung cells to ozone (Fig 3).

In cultured human diploid fibroblasts, morphological changes and

a gradual decrease in rate of growth have been attributed to a

buildup of cellular damage with each successive division

(11,12). Ozone may accelerate processes similar to those

naturally causing cellular damage and may decrease the growth

rate of the aging fibroblast colony. However, in ozone at 0.5

ppm, all of the human cancer cells (which do not age) had growth

rates several times lower than that of the aged, noncancerous

cells (Fig 2.).

Evidently, cancer cells are less able to compensate for the

oxidative burden of ozone than normal cells. The marked

sensitivity of cancer cells to ozone raises questions about the

possible mechanisms of oxidative inhibition of their growth.

Virtually every major component of normal cells has been found

to be affected by elevated ozone levels (5). However,

glutathione in its reduced form (GSH) has been credited with

providing the first line of defense against the peroxides and

free radicals generated in all cells by ozone and oxygen (1, 13-

15). It deactivates peroxides and radicals by donating one

hydrogen atom to the reactive species. Loss of a GSH hydrogen

(oxidation) results in formation of oxidized glutathione (GS-

SG). The cellular respiratory system is responsible for

reducing GS-SG to GSH. The GSH-linked respiratory system in

normal and cancer cells, before and after exposure to ozone,

must be examined to learn whether a functional impairment of

this system is associated with the marked sensitivity of cancer

cells to the oxidant.

These findings lead us to believe that ozone--alone, in

combination with radiation therapy (16), or in chemotherapy

utilizing electrophilic compounds (17)--may have therapeutic

value for patients with certain forms of lung cancer.

Frederick Sweet

Ming-Shian Kao

Song-Chiau D. Lee

Department of Obstetrics and Gynecology, Washington University

School of Medicine, St. Louis, Missouri, 633110.

Will L. Hagar

City of St. Louis Air Pollution Control, St. Louis, 63103

Wileen E. Sweet

Air Quality Section, East-West Gateway Coordinating Council, St.

Louis, 63102.

Figure 1. Schematic diagram (not shown) of the system used for

culturing human cells in ozonated ambient air. Filtered ambient

air was mixed with carbon dioxide (5 percent) and introduced

into a dual chamber incubator (National 331). Half was

conducted through a calibrated ozone generator consisting of a

quartz glass tube irradiated with ultraviolet light and then

into a hermetically sealed (20 by 20 by 20 cm) glass and

stainless steel environmental chamber containing a gasketed

access door. Output of ozone from the generator varied less

than 1 percent per day. The ozone content of the vented air

from the chamber was measured daily with a spectrophotometric

ozone analyzer (Dasibi 1003-AH). Malignant and normal human

cells were incubated in chamber E saturated with water vapor.

Corresponding cells serving as controls were incubated in the

adjoining compartment, also saturated with water vapor.

Figure 2. Inhibition by ozone of growth of malignant and non-

malignant cells in culture on day 8. Each of the cell types

were grown in 10 ml of Dulbecco's modified Eagle's minimum

essential medium containing 10 percent calf serum. In a typical

experiment, 12 dishes per cell line (usually three or four cell

lines were tested per experiment) were loaded into the

environmental chamber with an equal number of control dishes in

the adjoining compartment (Fig. 1). The initial population was

3 x 10(5) cells per dish. Every 48 hours three dishes for each

cell type were removed from both compartments and the cells were

tested for viability with 0.4 percent trypan blue and counted

with a hemocytometer. Each data point represents the number of

experimental cells divided by the number of corresponding

control cells per dish multiplied by 100 (the percentage of

control growth) and is plotted against the measured level of

ozone in the environmental chamber. The percentage of growth

inhibition is calculated by subtracting the percentage of growth

from 100. The data are from cell counting on day 8 of

incubation. There is a nearly linear relation between

inhibition of the growth of the cancer cells and increasing

ozone levels. The noncancerous cell line IMR-90 began to

display measurable growth inhibition only when ozone levels

exceeded 0.5 ppm, a level that produced approximately 60 percent

inhibition in all of the cancer cells lines tested. There was

some growth inhibition in noncancerous cells aged through 14

passages. The mean populations of the cells serving as controls

were as follows (per dish on day 8): IMR-90, 34.8 x 10(5); A-

549, 36.5 x 10(5); MCF-7, 57.0 x 10(5); endometrial

adenocarcinoma, 64.2 x 10(5); myometrial carcinosarcoma, 121.1 x

10(5).

References:

1. D.L. Dunsworth, C.E. Cross, J.R. Gillespie, C.G. Plopper, in

Ozone Chemistry and Technology. J.S. and J.R. Orr,

Eds. (lin Institute, Philadelphia, 1975), chap. 2.

2. H.E. Stokinger and D. Coffin, in Air Pollution, A.C. Stern,

Ed. (Academic Press, New York, 1968), vol. 1, pp. 446-546.

3. H.D. Kerr et al., Am. Rev. Respir. Dis. 111, 763 (1975).

4. J.D. Hackney, W.S. Linn, C.D. Law, S.K. Karuza, Greenberg,

R.D. Buckley, E.E. Pedersen, Arch. Environ. Health 30, 385

(1975).

5. B.D. Goldstein, Rev. Environ. Health 2, 177 (1977).

6. Normal human subjects tolerated breathing 0.5 ppm ozone in

air 2 hours per day for 1 week or 0.25 ppm ozone 2 hours per

day for 3 weeks (4). The two groups engaged in light

exercise during exposure. Although both groups developed

chest discomfort and moderately decreased respiratory

function during exposure, their removal from the oxidative

environment resulted in rapid disappearance of the symptoms.

The mean dose-response curves from this study show a no-

detectable-effect level at 0.25 to 0.30 ppm. A similar

study (3) found that human subjects tolerated exposure to

0.5 ppm ozone for up to 6 hours. Pulmonary function was

affected and chest discomfort developed at this level, with

no significant differences observed between smokers and

nonsmokers.

7. These cells (IMR-90) were obtained from the Human Aging Cell

Repository and plated 48 hours after shipping. This cell

type was characterized by W.W. Nichols, D.G. , V.I.

Cristofalo, L.H. Toji, A.E. Greene, and S.A. Dwight [science

196, 60 (1977)].

8. This cell line (A-549) was described by D.J. Glard, S.A.

son, G.J. Todard, P. Arnstein, J.H. Kersey, H. Dorsik,

and W.P. Parks [J. Natl. Cancer Inst. 51, 1417 (1973)]; M.

Lieber, B. , A. Szakal, W. -Rees, and G.A.

Todardo [int. J. Cancer 17, 62 (1976)]; and K.L. , N.S.

III, and J. [Cancer Res. 38, 1688 (1978)]

9. This cell line (MCF-7, estrogen-sensitive) was described by

H.D. Soule, J. Vazquez, A. Long, S. Albert, and M. Brennam

[J. Natl. Cancer Inst. 51, 1409 (1973)] and by K.B. Horwitz,

M.E. Kostlow, and W.I. McGuire [steroids 26, 785(1975)].

10. Human uterine carcinosarcoma cells and endometrial

adenocarcinoma cells were obtained from pathologically

confirmed gynecologic tumors and developed as new cell

lines. The endometrial adenocarcinoma cell line is

estrogen-sensitive. Both were described by M.S. Kao and

S.C.D. Lee (27th Annual Meeting of the Society for

Gynecologic Investigation, Denver, 20 to 23 March 1980),

abstr. 7.

11. J.R. and R.G. Whitney, Science 207, 82 (1980); S.C.D.

Lee, P.M. Bemiller, J.N. Bemiller, A.J. Papelis, Mech.

Ageing Dev. 7, 417 (1978).

12. P.M. Bemiller and L.H. Lee, ibid. 8, 417 (1978)

13. C.K. Chow and A.L. Tappel, Lipids 1, 518 (1972).

14. C.K. Chow, Nature (London) 260, 721 (1976).

15. R.E. Kimball et al., Am. J. Physiol. 230, 1425 (1976).

16. R.E. Lee, Semin. Oncol. 1, 254 (1974)

17. O.S. Selawry, ibid., p. 259.

18. Parts of this report were presented at the 27th Annual

Meeting of the Society for Gynecologic Investigation,

Denver, 20 to 23 March 1980 (abstracts 7 and 150), and at

the 73rd Annual Meeting of the Air Pollution Control

Association, Montreal, 24 June 1980 (poster session 27). We

thank W. -Rees for his gift of A-549 cells; the MCF-7

cells were obtained from E.M. Jensen. We also thank C.M.

Copley, Jr., H.M. Camel, and T. for their

constructive criticism of the manuscript. Correspondence

should be addressed to F.S.

24 April 1980; revised 11 June 1980.

moonbeam