The Mechanism And Influence Of Nasopharyngeal Cancer Stem Cell SP18Growth Affected By Carboxy Terminal Activating Region-3of The Epstein-barr Virus Encoded Latent Membrane Protein1

Objective: The pathogenesy of nasopharyngeal carcinoma is closely related withEpstein Barr-virus (EBV) infection. The latent membrane protein1(LMP1) wasknown for important oncogenic protein coded by EBV. This study aimed to observethe mechanism and influence of Nasopharyngeal Cancer Stem Cell SP18Growthaffected by carboxy terminal activating region-3of the Epstein-Barr virus encodedlatent membrane protein1and provided the experiment datas to reveal tumorigenesismechanism of EBV.Methods: Retrovirus RV-LMP1and RV-LMP1△232-351previously were collected toinfect the nasopharyngeal cancer stem cell SP18, respectively. The infected cells wereselected by geneticin, and then the cellular clones were collected. The SP18-LMP1and SP18-LMP1△232-351cell lines, stable expressed LMP1and LMP1of deletedcarboxy terminal activating region-3(LMP1△232-351) protein, were establish. Thegrowth of SP18cells affected by LMP1△232-351was detected by cellular growth curve,colony formation, and soft agar formation. The cell cycle was checked by flowcytometry (FCM), and then the cellular proliferation index was calculation. Moreover,the differential expression genes related with SP18cellular proliferation were detectedby gene chips. Late, the expression of part gene was measured by RT-PCR.Results:1.The result of cellular growth curve, colony formation and soft agarformation showed that, Compared with SP18-LMP1cell, the growth velocity and thenumber of colonies of SP18-LMP1△232-351cell were smaller and fewer than that ofSP18-LMP1(n=3，p<0.01).2. The result of flow cytometry (FCM) indicated that theproliferation index of SP18-LMP1△232-351cell was decreased than of SP18-LMP1cell (n=3，p<0.01).3. The genic chip all detected428differential expression genes, whichthey included those135differential expression genes that related with cellularproliferation (59genes up-regulation expression and76genes down-regualtionexpression).6. The expression of part gene was verified by RT-PCR as same as theresults of genic chip detection.Conclusion:1. The carboxy terminal activating region-3of LMP1was important activity site topromot SP18cellular proliferation.2. LMP1-CTAR3probably increased the cellular proliferation index to affect theproliferation of SP18cell.3. The carboxy terminal activating region-3of LMP1probably affected the expressionof IL1A and Wnt6genes to regulate the growth of SP18cell.