Tim Corcoran

Charlton Hilliard

7/26/10

BSC2010C

Cannabinoid Inhibit Cellular Respiration of Human Oral Cancer Cells

The scientists that preformed this study all have an extensive background in medical

research coming from all around the world. A few of them have actually worked together before

on similar studies. Harvey Penefsky and Abdul-Kader Souid had previously corroborated with

one another on a study of Doxorubicin and its effect on cellular respiration in mitochondria.

Their results from the research showed “cells incubated with Doxorubicin exhibit caspase-

mediated inhibition of oxidative phosphorylation.” So they have experience when dealing with

cellular respiration of cancer cells.

The intriguing part of this study is the use of a specific form of cancer cells, Tu-183,

which over expresses bcl-2(B-cell lymphoma 2) an apoptosis regulating protein that is highly

resistant to anticancer drugs. Thus the previous drug being researched, Doxorubicin, had little to

no effect on this strain. ∆9-tetrahydrocannabinol (∆9-THC) and ∆8-tetrahydrocannabinol (∆8-

THC) are known previously to significantly disturb the mitochondrial function and have

antitumor qualities. These compounds were then put to similar tests as Doxorubicin to fully

understand their effects on the mitochondria of Tu-183 oral cancer cells.

The study was preformed first by acquiring the certain reagents necessary for the

experiment. ∆9-THC and ∆8-THC were provided by the National Institute of Drug Abuse
(NIDA) and the endocannabinoid Anandamide was purchased from Biomol International. The

endocannabinoid (a compound similar to THC that our bodies produce naturally) was

instrumented to prove that the findings of this experiment were not due to the mediation of the

Cannabinoid receptors. The Tu-183 cells were acquired from State University of New York,

Upstate Medical University from Professor Edward J. Shillitoe.

The samples of ∆9-THC and ∆8-THC were then tested against the two laboratory standard

samples using high-performance liquid chromatography. There were found to be identical to one

another and thus enabling the continuation of the experiment. The cellular respiration of Tu-183

was then measured to provide a basis to compare the results from the addition of ∆9-THC and ∆8-

THC. Mitochondria extracted from beef heart were then suspended and induced respiration with

the introduction of succinate. Afterwards a control followed by ∆9-THC was introduced to show

the decline in the rate of respiration. The amounts of ∆9-THC and ∆8-THC were then varied to

determine concentration dependence. Cellular ATP was also measured in the Tu-183 cells.

Finally DNA fragmentation was utilized through the TUNEL technique to show the physical

effects of the administration of ∆9-THC.

The results of the first experiment were done at increments of differing amounts of ∆9-

THC and ∆8-THC with an increased inhibition of cell respiration with the increase in amount of

∆9-THC and ∆8-THC. The O2 consumption for untreated cells was 1.7 µmol/lO2min-1. At 10

µmol of ∆9-THC the consumption was 1.9 µmol/lO2min-1 (insignificant result). At 20 µmol of ∆9-

THC the consumption was 1.7 µmol/lO2min-1. At 50 µmol of ∆9-THC the consumption was 1.0

µmol/lO2min-1(41% inhibition). At 100 µmol of ∆9-THC the consumption was 0.8 µmol/lO2min-
1
(53% inhibition). The same experiment was then replicated and the O2 consumption for the

untreated cells was 1.9 µmol/lO2min-1. At 10 µmol of ∆8-THC the consumption was 2.0
µmol/lO2min-1 (insignificant result). At 20 µmol of ∆8-THC the consumption was 2.0

µmol/lO2min-1(insignificant result). At 50 µmol of ∆8-THC the consumption was 1.8

µmol/lO2min-1. At 100 µmol of ∆8-THC the consumption was 1.3 µmol/lO2min-1(32%

inhibition).

The following experiment determined the exact time of the onset of the effects of ∆9-

THC and ∆8-THC by testing the treated cells at different intervals. The results determined that

the effect is immediate. This adds to the devastating effects found in the last experiment proving

its worth as a medicinal plant.

The beneficial effects determined by this experiment have such an extreme potential for

the health of mankind. Being a plant anyone with illness can just place a seed in the ground,

give it water and wait for it to bear fruit. This also means that once legalized businesses can

produce it extraordinarily cheap compared to the prices of alternative medicines. This wide

availability would do wonders for people who cannot afford the cost of health care.

The history of cannabis has been negative to say the least; this present information may

inspire an honest and thorough investigation into all of the plant’s positive benefits. Although

the prejudice held currently by many people today regard marijuana as a gateway towards

harmful chemicals, but I see no difference between that effect from marijuana and alcohol.

Substitution of cannabis with alcohol as a recreational drug would help those seeking alternative

states of mind or realities.

The side effects of alcohol consumption as most people are aware of is not only similar to

poison towards the body, but also leads to making terrible decisions that could endanger one’s

self and those around him. Cannabis on the other hand produces a calm, soothing effect that not

only leaves one clear to think properly, but also wears off relatively quick in comparison with
alcohol. These wildly known effects are a commonly sought enjoyment by a growing number of

people not only in United State of America but also many other countries around the world. This

is obviously only one of the many benefits that this plant gives to the human race, including the

power to heal.

With marijuana’s active ingredients having the ability to slow cell respiration in cancer

cells one is left to wonder the effects of the “War on Drugs” against it. Take for example a nice

older gentleman named Rick Simpson. Mr. Simpson has been creating hemp oil from cannabis

plants and successfully cured patients of his from many illnesses. He states, “The results of the

cases can be readily replicated by any practitioner, medical or otherwise, anywhere, to cure

malignant melanomas and more importantly, save lives. The topical application of hemp oil

salves or balms helps to control or cure various skin conditions. Taken orally, the oil tends to

seek out and destroy cancer cells in the body”.

When Rick Simpson then won the Freedom Fighter of the Year 2009 at the Cannabis Cup

in Amsterdam his house in Canada was then raided and he was instructed that if he ever came

home he would be arrested. His site boasts pages of accounts of people curing their diseases

from using hemp oil and all the powers that be can think is to put him in jail. He explains our

current legal situation with cannabis as, “The current restrictions against hemp were put in place

and maintained, not because hemp is evil or harmful, but for big money to make more big

money, while we suffer and die needlessly. Look at a proposal such as this; if we were allowed

to grow hemp in our back yards and cure our own illnesses, what do you think the reaction of the

pharmaceutical industry would be to such a plan? Many large pharmaceutical companies that

still exist today sold hemp based medicines in the 1800's and early 1900's. They knew then what
I have recently found out. Hemp oil if produced properly is a cure-all that the pharmaceutical

industry can't patent.”

Although Mr. Simpson has extensive experience with cannabis he does note that he does

not have a medical degree. The medicinal properties of cannabis that Mr. Simpson is attempting

to bring to light have been utilized for thousands of years by our ancestors. It does not take an

eight year degree to determine whether a four month treatment of cannabis oil prior to a CT scan

showing no spreading of the cancer as well as outer layers of cancer dying. Then one year from

the start of the cannabis oil treatment this patient’s cancer had been cured. Of course these

people cannot come forward and explain their findings due to the legality of the issue. This

binding legality has kept some of the wonders of nature under wraps from the people of today.

Another positive aspect of cannabis cultivation and use is the fibers from the stems of the

plant itself and its relative hemp (a strain with 0% THC) can be used to make products

commonly made with cotton. According to a book written by Chris Conrad he notes the higher

yields that can be produced from cannabis cultivation. “An acre of land will produce about 1000

pounds of primary hemp fiber, about 2 or 3 more times fiber than cotton. Fiber comes right off

the plant ready to comb and use.” This man also goes on to talk about the history of its use

throughout the time of man back to the Rig-Veda, a traditional Hindu story dating back about

3500 years ago.

The fibers of the cannabis plant can also be used to make paper and help slow the

destruction of rainforests and homes of animals. A crop of cannabis can be cultivated in 100

days giving it an extraordinary capability of producing what we need if the legality were

different. The so called “drugs”, mainly just the natural, of our time are scary, but their

beneficial properties cannot be ignored any longer. The same government that enabled us as a
society to grow to the state we are in today has an obligation to recognize the fallacies of our

current policy and devise a new plan or measure in ending modern prohibition and the utilization

of the natural wonders given to us by the loving planet that we inhabit.

Progress towards the truth and future of cannabis within our culture has taken many of

the necessary steps in not only giving people the medicine that they deserve, but saving the

innocent people currently being prosecuted by pointless and harmful laws banning a plant. 19

states currently have laws that lower the limits set by the federal government. Proposition 19

hopes to legalize cannabis use in California in the 2010 November ballot. It would let people 21

and older to cultivate, use and travel with designated amounts of cannabis. This measure with

also probably give way to taxation of the plant and give extra needed money to our already

struggling economy.

Some states have dropped the laws of cannabis to a simple ticket instead of a

misdemeanor, otherwise known as decriminalization, or they have recognized it medical benefits

and allowed people to obtain licenses to cultivate and/or use the plant medicinally. These

measures are certainly beneficial to the people that they help and the local governments receive

needed tax dollars, but a much less limited system is necessary to rid all the injustice that plagues

our modern society.

The National Organization for the Reform of Marijuana Laws (NORML) have a

wonderful website showcasing the latest news and updates in the topic of marijuana as a

medicine, a recreational herb and its legal status in every part of the United States. This study

was posted on its website and distributed to its email readers, giving people the truth about

cannabis and its benefits. Without NORML’s formation in 1970 much of what we have

accomplished now may have never been realized. Their struggle has a been rough and extensive,
but the battles remain to be fought and with the help of donations they use every tool of modern

society to give knowledge to the people of the world.

What this study has produced is clear and concise evidence of what the people of

NORML, Rick Simpson and many other intelligent people have known for years. Although it

has not given absolute answers to the many questions surrounding cannabis, but it does leave a

path that needs to be traversed fully. Once this path has been followed, documented and shared I

feel the knowledge derived from such finding will change our society and way of life more than

people can anticipate or fathom.

 References

Conrad, C. (1994). Hemp: lifeline to the future. Creative Xpressions.

Marijuana law reform. (n.d.). Retrieved from http://norml.org/index.cfm

Whyte, D.A., Al-Hammadi, S., Balhaj, G., Brown, O.M., & Penefsky, H.S. (2010).

Cannabinoids inhibit cellular respiration of human oral cancer cells. Pharmacology, (85),

Retrieved from http://www.karger.com