| Objective: We injected BALB/c A-nu nude mice(female, 4 weeks old) with Human Hepatocellular Carcinoma Cell Lines Hep G2 and Li-7, respectively, to observe whether they can cause oncogensis. And then we detected the the Serum Proteome Profiles of nude mice in different times, using the Surface Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry(SELDI-TOF-MS) to screen the proteins related to liver cancer. Method: We cultivated the 2 cell lines and selected the ones in logarithmic phase to subcutaneously inject them into the left upper limb of nude mice in the back side with the same dosage. The control groups were injected with RPMI-1640 Medium. We observed them regularly and weighed them every week. We collected the whole blood from eyes. For the mice injected with Hep G2, their blood was got at the 20 th, 40 th, 60 th day from the day they were injected, including the control groups. For the mice injected with Li-7, their blood was got at the 40 th day, including the control group. We also harvested their livers, lungs, kidneys and stomachs for histopathology. After the whole blood clotted, they were centrifuged for 6 minutes, with a revolving speed of 6000 revolutions per minute. We put the serum in new eppendorfs and reserved them in the ultra-low temperature(-80 ℃) freezer. Then, we analyzed the proteome profiles, using the inherent softwares(Ciphergen Protein Chip 3.0 and Ciphergen Biomaker Wizard 3.1) in SELDI-TOF-MS and SPSS 16.0, and got the differential proteins. Results:(1) The 2 Human Hepatocellular Carcinoma Cell Line Hep G2 and Li-7 cannot cause oncogenesis. The viscera of mice injected cell line Hep G2 are not abnormal. The liver is oedematous and the stomach is focal intestinal metaplasia in the mice injected cell line Li-7.(2) There are 9 differential proteins in the control groups of mice injected cell line Hep G2 and 3(M/Z is 4014.16, 4122.70 and 8373.05) of them are upregulated and 3(M/Z is 1264.62, 1491.80 and 2582.5) of them are downregulated. There are 29 differential proteins in the treatment groups and 5(M/Z is 3701.15, 4560.87, 8169.14, 8258.08 and 12047.4) of them are up-regulated and 3(M/Z is 1025.35, 1045.15 and 4319.54) of them are down-regulated. These regularly varied proteins cannot be observed in the control groups. There are 4 shared differential proteins(M/Z is1264.62, 1491.80, 4014.16 and 8373.05) in both control and treatment groups. In the control groups, proteins of 4014.16 and 8373.05 are upregulated and 1264.62 and 1491.80 are down-regulated, but in the treatment groups, all the 4 proteins vary irregularly.(3) In the mice injected cell line Li-7, there are 12 differential proteins(M/Z is 1027.39, 1087.44, 1179.59, 1192.62, 1483.17, 1563.20, 3763.00, 3869.26, 4319.54, 6646.16, 8793.72 and 9259.75). Conclusion: 1. Human Hepatocellular Carcinoma Cell Line Hep G2 and Li-7 are not oncogenous. Not every kind of Liver Cancer Cell Line can cause oncogenesis in nude mice. 2. SELDI-TOF-MS detects the proteome profiles and screens the differential proteins, proving a new orientation of diagnosis and research in liver cancer. 3. The types and quantities of proteins are different in the serum of mice injected different liver cancer cell lines, suggesting that every liver cancer cell line has its own exclusive proteins. |
PDF Full Text Request