I need the most common methods and procedures to screen a phytochemical having anticancer activity both in vivo (animal model) in vitro and in computational systems.
For invitro you can use MTT assay, it is quite sensitive and will help you determine the IC50
1. Cell Culture dependent cytotoxic screening [Normal mouse fibroblast cells (NIH/3T3,ATCC: CRL-1658) and three human cancer cell lines gastric adenocarcinoma cells (AGS, ATCC: CRL-1739), colorectal adenocarcinoma cells (HT-29, ATCC: HTB-38) and breast ductal carcinoma cells (MDA-MB-435S, ATCC: HTB-129) can be used for cytotoxicity screening]
2. MTT assay: The cytotoxicity of the extracts will be tested against B-16 (murune melanoma), HCT-8 (human colon carcinoma), CEM and HL-60 (leukemia) tumor cell lines.
Brine shrimp lethality may eacy way to identify either the extract toxic to cell.
You can also do proof-of-efficacy assay depending upon mechanism of action e.g. antioxidant activity (in animal skin) or angiogenesis inhibition (in egg/chicken egg).
A good starting place will be in the heart of the pit of the aprocott seed where the Vitamin B-17 comes from that makes 2 enzymes in the human body - 1 unlocking in the cancerious part of the body - and 2 a relocking enzyme in the healthy part of the body orenting themselves respectively via pH. In the Vitamin B-12 the cobalimin gets unlocked from the cyanide that kills the cancer - then hits the relocking enzyme that relocks the cyanide into endorphrens and secondary pain killers. The liers in the AMA and the FDA (pupet organizations for the pharmaceuticles) have tried to turn this country into a third world medical country and worked very hard against the American people. Good Luck in this worldwide fight against stupidity that will eventually be against the establishment medical comunity.
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