Experimental Study About The Effect Of Chemokine Receptor In T-cell Non-Hodgkin's Lymphoma Dissemination

ObjectiveThe expression of chemokine receptors CCR7 and CXCR4 has been associated with tumor dissemination and poor prognosis in a limited number of tumor entities. However, there is not so much data currently available on the impact of chemokine receptor expression on disease progression and prognosis in human T-cell non-Hodgkin's lymphoma(T-NHL).This is a pilot study about the role and mechanism of chemokine receptors and some other cytokines in T-NHL invasion process.MethodsPARTI:Specimens of 41 T-NHL cases and 19 reactive lymphoid hyperplasia cases were collected along with their clinicopathologic data. The expressions of CCR7, CXCR4, MMP-2, MMP-9 and VEGF were detected using immunohistochemical staining and were then used to study the correlation between the expression and clinicopathologic factors.PART2:The human T-NHL cell lines Hut78 (cutanous T-cell lymphoma) and Jurkat (adult T-cell leukemia/lymphoma) were cultured, then the transcription and expression of CCR7, CXCR4 and key enzymes in PI3K/Akt pathway were evaluated using RT-PCR and Western Blotting. The invasion ability of the two cell lines were measured by Transwell invasion assay and then used to study the impact of chemokine receptors in T-NHL invasion process and the molecular mechanism that may involved.Result and ConclusionPARTI1. The expression of CCR7, CXCR4, MMP-9, MMP-2 and VEGF were significantly higher in T-NHL group than in the control group. Overexpression of CCR7, CXCR4, MMP-2, MMP-9 and VEGF were detected in T-NHL.2. The overexpression rates of CCR7 and MMP-9 were higher in patients with multiple lesions than those in patients with single lesion. And strong expression of MMP-9 was significantly associated with higher stages 3/4 and larger size of tumors(>3cm).In contrast to MMP-9, the overexpression rate of MMP-2 was higher in cases with single lesion than in cases with multiple lesions. CCR7 and MMP-9 may be the unfavorable factors influencing prognosis of T-NHL while MMP-2 may be a favorable one.3. The positive correlation between the expression of CCR7 and CXCR4, CXCR4 and MMP-9 were showed. These molecules can be considered as predictors of outcome individually or collectively.PART21. The transcription of CCR7, CXCR4, PI3K and Akt was significantly stronger in T-NHL cell lines Hut78 than in T-NHL cell lines Jurkat, the expression of CCR7, Akt and p-Akt were correlated with the respective transcription profile. And Hut78 cell line showed stronger invasive ability in Transwell invasion assay. The transcription and expression of CCR7 and CXCR4 may play an important role in the invasion process in vitro in T-NHL. And the activation of PI3K/Akt signal transduction pathway may be involved as a down-stream pathway.2. The T-NHL cell lines have been co-cultured with chemokine CCL21, and then: Both Hut78 and Jurkat showed stronger invasive ability than the respective control group. CCL21 may enhance the invasive ability of T-NHL cell lines. And Hut78 showed stronger invasive ability, higher level of transcription and expression than Jurkat before and after the co-culture with CCL21.3. After co-cultured, the transcription of CCR7, CXCR4, PI3K and Akt, the expression of CCR7, Akt and p-Akt were up-regulated. Increased invasive ability and up-regulation of the transcription and expression in two cell lines are mainly correlated with the concentration of CCL21. The fact that the transcription and expression of CCR7 and CXCR4 may impact the invasion process in vitro in T-NHL and the activation of PI3K/Akt signal transduction pathway may be involved as a down-stream pathway was further proved.