| Liver cancer which has a very high morbidity in China has seriously menaced the healthy of Chinese people.Nowadays,to deal with liver cancer which can not be removed by operation or to prevent the recrudescence of liver cancer which has already been removed by operation,chemotherapy is still the main methods of treatment.Thalidomide THD,one kind of Glutamic acid derivatives which was used to treat Pregnancy vomiting,but has finally been removed from the market because it has the teratogenic effects in the 1960s,has been discovered to be effective in some treatment for some autoimmune diseases.Recent research reveals that THD is a potential antitumor drug.It can play the role of liver cancer through inhibiting new blood vessels produce,regulating the immune function,etc.5-fluorouracil,a classical anti-metabolism type of cancer chemotherapy drug which has been used to deal with sarcomata,is still the main drug to treat primary liver cancer.But it has some disadvantage such as the heavy side effects,the short half-life, drug-resistant,etc.This assay will discuss the effect and principle of treatment for liver cancer by using Thalidomide in combination with 5-fluorouracil through experiment inside and outside body.Points are as follows.1.The effect Thalidomide in combination with 5-fluorouraeil played in defending liver cancer outside bodyWe detect the inhibition played by thalidomide in combination with 5-fluorouracil on liver cancer cell cultured in vitro using MTT.Thalidomide in combination with 5-fluorouracil in the dose of(6.25 +3.125) -(100 +50)μmol L-1 has inhibition on both mice HepA liver cancer cell lines cultured in vitro and human HepG2 liver cancer cell lines cultured in vitro.The higher concentration is,the stronger effect has,with has a clear dose-effect relationship.The inhibitory effect of different cell lines is different,the IC50 values are:[28.9μmol L-1(THD),14.4μmol L-1(5-Fu)],[33.9μmol L-1(THD) and 17.0μmol L-1(5-Fu)],they are all stronger than single-use thalidomide in(73.2, 78.6μmol L-1 and the single-use 5 - Fu(29.2,34.9μmol L-1),and the two drugs have synergies.Treat with HepA and HepG2 respectively by using thalidomide in combination with 5-fluorouracil(50 +25)μmol L-1 and observe the growth of cell after 12,24,36,48 hours respectively,we finally find that the effect of Thalidomide in combination with 5-fluorouracil on tumor cells has an obvious time-response relationship,the longer time,the stronger effect.After 24-48 hours,using thalidomide in combination 5-fluorouracil has a stronger inhibition than by using the same dose of thalidomide or 5-fluorouracil.2.The effect Thalidomide in combination with 5-fluorouracil played in defending fiver cancer inside bodyThe administration of Thalidomide in combination with 5-fluorouracil for 13 consecutive days has the significantly inhibition in the growth of the HepA hepatocellular carcinoma tumor inside the mice.The inhibition rates of Thalidomide in combination with 5-fluorouracil[(20 +20),(40 +20)(80 +20) mg·kg-1×13d]were 38.2,56.2,66.1%,which is significantly higher than the rates of using the same dose of thalidomide or 5-fluorouracil alone.Besides,the use of Thalidomide in combination with 5-fluorouracil can significantly prolonged the survival time of HepA hepatocellular carcinoma model mice,the survival extended rates of Thalidomide in combination with 5-fluorouracil[(20 +20),(40 +20)(80 +20) mg·kg-1]is 35.5,80.8,129.6%respectively.3.The side effects Thalidomide in combination with 5-fluorouracil played in defending liver cancer inside bodyThe use of Thalidomide in combination with 5-fluorouracil[(20 +20),(40 +20)(80 +20) mg·kg-1×13d]significantly increased the speed of growth of HepA liver cancer mice's weight,and the effect is stronger than that of using 5-fluorouracil 20 mg·kg-1. The use of Thalidomide in combination with 5-fluorouracil(80 +20) mg·kg-1 can significantly improve the reduction of leukocyte count in the 5-fluorouracil 20 mg·kg-1 group.In addition,the use of Thalidomide in combination with 5-fluorouracil(40 +20) mg·kg-1,(80 +20) mg·kg-1 significantly increased the spleen index of HepA liver cancer mice,and the effect is stronger than that in 5-Fu 20 mg·kg-1 group.4.The research on the Anti-hepatoma of Thalidomide in combination with 5-fluorouracilThalidomide in combination with 5-fluorouracil(25+12.5)μmol L-1,(100+50)μmol L-1 can evidently restrain TS outside body and the effect of inhibition Thalidomide in combination with 5-fluorouracil(100+50)μmol L-1 played is heavier than the effect of inhibition single 5-fluorouracil with the same dose.The HepG2 cells with the treatment by the thalidomide in combination with 5-fluorouracil show typical changes in tumor cell apoptosis behind electron microscopy. Apoptotic cells of which the volume is reducing can be seen that their chromatin condense to dense and distribution become a crescent-shaped along the nuclear membrane.Advanced apoptosis cell can be seen that the cells decompose into apoptotic bodies coated whir the cell membrane.With DNA labeling,flow cytometry make analysis of apoptosis change.While Thalidomide in combination 5-fluorouracil(6.25 +3.125) -(100 +50)μtool L-1 act on HepG2 for 48 hours,apoptosis rate rise from 10.9%up to 37.8%percent,showing a significant concentration-dependent,and the apoptosis rate of combination(100 +50)μmol L-1 is higher than that of the same dose of 5-fluorouracil.Thalidomide in combination with 5-fluorouracil(40 +20),(80 +20) mg·kg-1 significantly increases the proliferation of spleen lymphocytes induced by HepA liver cancer mice ConA,LPS,and the effect is obviously stronger than that using 5-fluorouracil 20 mg·kg-1 only.In addition,Thalidomide in combination with 5-fluorouracil(40 +20),(80 +20) mg·kg-1 significantly increases the production of IL-2 and the activity of NK cell inside liver cancer mice,inhibit the abnormally high content of NO inside HepA liver cancer mice,and the effect is obviously stronger than that using 5-fluorouraci120 mg·kg-1 only.5.ConclusionThalidomide in combination with 5-fluorouracil has an evident effect in defending the liver cancer both inside and outside body and the effect is stronger than that of using Thalidomide or 5-fluorouraeil only.The mechanism may be related with inhibiting the express of TS and increasing the withering of the tumor cell,accommodating the function of the immunity.
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