الفهرس 
Introduction
CancerOnly 14 pages are availabe for public view
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Abstract
Despite the tremendous medical boom witnessed by the world in the treatment of cancer, but the science is so helpless in front of an effective drug is devoid of side effects to alleviate this deadly disease that spreads among humans rapidly attacks .. With the passage of time doctors realized the seriousness of what they’re using from chemical treatments, radiological and immunological and genetic and other ...... sick, recently opted their eyes to the unexpected ways to complete the permanent war against cancer.
Most countries of the world and the suffering that all of them were not from the spread of cancer diseases which pose a serious threat to both human element of the national economy over the ages and ages, and in all developed and developing countries of the world , not least because of the statistic in 2012 , which was published by WHO global in 2014 that cancer of the most important causes of morbidity and mortality throughout the world, including approximately 14 million new cases and 8.2 million deaths related to cancer in 2012 only colorful forms as follows:
• Lung cancer (1.59 million deaths).
• Liver cancer (745.000 deaths).
• Stomach cancer (723.000 deaths).
• Colorectal cancer (694.000 deaths).
• Breast cancer (521.000 deaths)
• Cervical cancer (400.000 deaths).
Despite strenuous efforts by developed countries to find an effective treatment for cancer, but they are still not able to find an effective treatment without risk.
US researchers have found through a series of laboratory tests that scorpion venom can now be counted on to deal with this severe disease dangerous.
The researchers explained that the scorpion sting launches a mixture of nerve poisons that contain peptide or protein associated with some cancer cells but not close to healthy cells , and it was French doctors had confirmed earlier that the scorpion is possible to perform the duty of solemn in the eradication of cancer cells from the body, because the needle in the guilt of the scorpion and the container able to poison Identified the remains of cancer cells very clearly after the addition of radioactive materials to the amount of venom injected into the site of infection , and thus can keep track of those parts and completely removed. There have also been attempts in the field of ancient folk medicine use Chinese scorpion venom in cancer treatment and found that the extract of scorpion venom dramatically affects the cancer cells to change shape and the loss of the outer layers of the cell cancerous , leading to constriction and cell death.
There are various types of cancer and is the most common lung cancer and there are several types of cancers such as cancers of the breast, liver, blood, stomach, colon, cervical and other........ Etc.”
Based on what has been the studies concern earlier this message was aimed at trying to study evaluating the use of scorpion venom as an effective means to eliminate the cancer cells, and to study the possibility of its use as a safetreatment and effective with no side effects.
At the beginning, scorpions are one of the types of arthropods toxic , one of the members of the spiders , and up the types of scorpions to 1,500 species , including 50 species of the most toxic in the world’s species, and spread all over the world except Antarctica and divided scorpions to 18 families are considered ”buthidae” most family killed and toxicity and the most medically important. As for the scorpion venom every type of scorpion venom has a different type, and consists of scorpion venom between 50-100 different type of protein toxic substances and a few dozens of species are well-studied.
Scorpio marking contains about 250 biologically active proteinsand peptides which considered as an anti -tumor and is under discussion. It causes by scorpion venom peptides, that disrupt the cell membrane which affects the flow of movement and leads to polarization fought those ions Vijlq pores Phones in the cell membrane than the cellular content leak and destroy the cell membrane cause which affects a number of vital functions within the cell, such as fat distribution and others.
In this study, we used scorpion venom (Leiurusquinquestriatus) and (combretastatin) which is used as a chemical therapy in the treatment of tumors to study the impact of the venom on the cancer cells (human breast cancer cells (MCF7) - liver cancer(HepG2) - cell lung cancer cells (A549) ) and compare with the results of combretastatin chemotherapy which act as the reference material,alsouse healthy cells to determine the impact each of scorpion venom and chemotherapy and we differentiate between treatments by both of themby using embryonic cells in human lung(MRC5)for this purpose ( healthy cells ) by conducted the following tests : -
1. Test (MTT).
2. Test (ELISA).
3. Test (PCR).
The observed results revealed the following:
We tried to prove the effectiveness of scorpion venom (Leiurusquinquestriatus) on some types of cancer cell lines as mammarian cancer cell line ”Human breast cancer cell line” (MCF7); liver carcinoma cell line (HepG2) and lung adenocarcinoma cell line (A549) and human fetal lung (MRC5) as a normal cell line which have been demonstrated in vitro in several analytical assays as MTT,TubulineELISA and PCR.
Initially we had made MTT assay (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) and we found a satisfactory results for all cancer cells that we have analyzed. By mean, we observed a dose- and time-dependent anti-proliferation effect of scorpion venom on these cells.
The scorpion venom -induced apoptosis might be mediated by the activation of intrinsic mitochondrial pathway and extrinsic death receptor pathway through regulation the expression of related apoptotic factors. The exposure of cancer cells to scorpion venom evidenced a significant cytotoxicity and selectivity against epithelial cancer cells while slight effect was observed on epithelial normal counterpart or no effect.
Among all our experimental epithelial cancer cells, We observed that in MTT cytotoxicity assay were had very good viability percentage detected by IC50 (concentration of venom that inhibits cell growth by 50%)values, the sensitivity of HepG2 (hepatocarcinoma cell lines) were the most one (IC50=0.11μM) followed by MCF7(breast cancer) which give satisfied result (IC50=0.15μM) , while A549 ( lung adenocarcinoma) were the least one (IC50=0.29μM) but consider satisfactory result, comparing by the reference (combrestatine chemotherapy) were give results with IC50 higher than all IC50 treated by scorpion venom Which indicates the venom has %viability better than the reference chemotherapy , With a vision that the effect of venom on the normal cells (MRC5)slightly toxic (IC50= 1700μM)which indicated that absence or weakness of scorpion venom to healthy tissues. Also we noticed that whenever the concentration of the venom increased,the more impact of it appeared in a better way which leads to the high proportion of %viability.
Additionally to MTT assay, we support our result by other investigations.One of them was ELISA assay (Tubulin enzyme assay) which indicated % Tubulin inhibition in every cell of A549 , HepG2 , MCF7cells according to its IC50 values were detected in MTT assay and we found that A549 , HepG2 cells were nearly parallel to each other (92.26% and 92.22% respectively) more than MCF7 (89.21%).
In general all our cells give results of tubulin inhibition better than Combretastatin, when we treat it on HepG2 cells give result (86.68%),while % Tubulin inhibition on untreated HepG2 by venom which act as a control, equal zero which indicate that the only sources which act as a poptotic marker on cancer cells were venom and Combretastatin with high proportion to venom than chemotherapy. Thus, induction of apoptosis in cancer cells is a key mechanism by which anticancer therapy works.
This study showed thequalitative technique as PCR assay measured some of optimal activities expression of main caspases related with the extrinsic and intrinsic apoptotic pathway in every cancer cell lines which treated (MCF7, A549, and HepG2).
Active forms of Capase-3, 8, 9 were detected within 40 cycles and less CT (cycle number) value mean faster affection by L. quinquestriatusvenom after treatment and higher mRNA of caspase expression, while in untreated control cells there was not observed increment of active form of cancer cell gene expressions. Active forms of Capases-3, 8, 9 gave us satisfied indications about apoptosis induction .it was detected bymultiple fluorescence signals which gave elevated threshold pathways are involved in the regulation of apoptosis.
The data of this study showed a significant effect of L. quinquestriatusscorpion venom against MCF7, A549 and HepG2 appeared in models depending upon CT value (cycle per time value) which detected by reaching to the threshold and we observed good expressions with threshold cycles of all our cancer cells which under our study (MCF7 , A549 , HepG2). We confirmed the validity of these result by positivity of GAPDH (β actine act as internal positive control: (25.16 cycle)) and undetectable result of negative control (water).
Graphics of CT values of scorpion venom against different cell lines which analyzed the three main caspases of every cancer cell (caspase 3, 8, 9) represented significantly over-expressed and accelerated in CT-dependent manner we saw that in the previous graphs of different elevations in threshold until reaching to plateau stage which no more products or reagent occur. So caspeses3, 8, 9 in MCF7 which treated byL. quinquestriatusscorpion venom gave CT values 25.83, 23.46, 26.93 cycles respectively.And the same previous CT values of HepG2 caspases were in the same order 29.38, 27.55, 25.05cycles. Finally A549 caspases which consequential as beforewere represented CT values 22.76, 21.05, 24.35cycles.
from the above draw this, when MCF7 , A549 and HepG2 treatedwith L. quinquestriatusscorpion venom, it give multiple significant CT values in all cancer cells which indicate the ability of scorpion venom to influence on pro-apoptotic genes and cause apoptosis and the evidence is faster CT values from the panel which correspond to A549, MCF7, HepG2 respectively.
from the previous results, we observe that A549give thebest CT values with its main caspases which illustrated to us that the L. quinquestriatus scorpion venom has significant impact ongeneti expressions of the proteolytic enzymesof lung adenocarcinoma. And also it able to influence in a satisfactory manner with MCF7 caspases CT values as evidence the ability to L. quinquestriatus scorpion venom effect on breast cancer. While HepG2caspases CT values gave resultsmust reckoned with it which confirm L. quinquestriatus scorpion venom ability to influencehepatcarcinoma. All apoptotic marker which detected, give us indication to ability of scorpion venom to enter the cancer cell in the cell programming of death.
The conclusion revealed that Venom from Leiurusquinquestriatusscorpion induces selective and differential anticancer effect against epithelial cancer cells. This study demonstrated that the apoptotic and necrotic effect of venom was probably due to the sensitivity of cancer cells; however, further studies involving more cell lines at different doses of natural products are needed to firmly establish these observations. To our knowledge this is the first scientific evidence demonstrating the effect on cell viability and cell death events of this scorpion venom in a panel of human cancer cell lines. This makes Leiurusquinquestriatusscorpion venom a favorable candidate for future studies in vivo anti-tumor experiments.