Hepatoma Cell Qy And Its Parvovirus H1-resistant Cells The Cloning Qyrc Gene Expression Differences

Autonomous parvovirus H1(PV H1) is a kind of nonenveloped,lytic DNA viruses which is cytotoxic to human and animal(mouse or dog) tumor cells caused by different inducing factors(DNA and RNA tumor virus or chemical carcinogenic agent) while innocuous to non-transformed cells.Some studies have found that the different sensitivity to PV H1 of tumor cells and their normal counterparts is due to their distinct gene expression.Although previous studies have identified dozens of cellular factors which are related to the parvovirus infection,it still remains lots of questions including which genes are involved into the process? How to explain the mechanism underlying?HepatoceUular carcinoma is considered as one of the most common cancers in China and the southern Asia.The interaction between human hepatoma carcinoma cell line QGY-7703 and PV H1 have been investigated in the previous works.In order to gain a insight into the difference at the transcriptional level between the PV H1 resistant ceils and PV H1 sensitive cells,QGY-7703 was considered as the parent cell, treated with PV H1,and acquired a couple of "counterpart cells"(including Parvovirus H-1 resistant cells and PV H1 sensitive cells).We adopted both the DNA microarray technology and the real-time PCR technology to investigate the global expression profiling differences of these "counterpart cells";hoping to discover those genes which are related to the parvovirus infection,and to threw some light on further cancer research.In this study,PV H1 sensitive cell QY was acquired from hepatoma carcinoma cell line QGY-7703 by screening a clone from soft agar QGY-7703 clones,while PV H1 resistant cell QYRC was acquired from QY by repeated PV H1 infection,and screening a single stable clone.The different biological characters of these "counterpart cells" are studied by cell biology and virology experiments,respectively. Then carrying out the oligonucleotide microarray experiments which represented 38, 500 humen genes(Genechip Human Genome U133plus2.0,Affymetrix) to study the gene expression profiling differences in the "counterpart cells".Finally,validate the expression differences of candidate genes chosen from the gene chip results by Real-time PCR.From the results,we found that,the biology character of PV H1 resistant cell QYRC,which had similar characters with normal cells was much different from PV H1 sensitive cell QY and QGY,which were hepatocellular carcinoma cells.And the gene chip results indicated that in the global mRNA expression level,compared with PV H1 sensitive cell QY,there is a larger number of up-regulated genes in PV H1 resistant cell QYRC.Combined with 2 previous gene chip results based on 2 different "counterpart cells"(L02C3RCR50 vs.L02;L02C3RC1 vs.L02C3),we at last acquired 19 co-up-regulated,significant express changed(SLR=1) genes and 1 co-down-regulated,significant express changed(SLR=1) gene.Then we proved the microarray results of 16 significantly changed(SLR=1) genes from cytology and clinical histology aspects,respectively,by SYBR Green Real-time PCR.The PCR results indicated that the express change of most genes coincided with microarray results.Thus the microarray results are proved reliable.Above all,our results suggested that there are more genes expressed in PV H1 resistant cells than PV H1 sensitive cells.The changed genes relates to many different biological process and pathway.With a large number of genes involved in signal transduction,transcription regulation,cellular adhesion,cell migration,inflammation,etc,the result indicates that the "counterpart cells" changed a lot in gene expression.Those identified genes could be related to the PV H1 infection and induction of cancers,which need further identification.