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Ren Y.-X.,Head and Neck Tumor Research Center | Li X.-J.,Head and Neck Tumor Research Center | Yang J.,Head and Neck Tumor Research Center | Ma J.,Kunming Childrens Hospital | And 6 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2012

Objective To investigate the expressions of BCRF-1mRNA and VIL-10 protein so as to clarify the effect of VIL-10 on immune cells of nasopharyngeal carcinoma (NPC). Methods The mRNA of BCRF-1 in tissues of NPC was detected by RT-PCR and VIL-10 protein was detected by Western blot. Flow cytometry was used to calculate the ratio of CD4 + T cells to CD8 + T cells. Results (1) The mRNA of BCRF-1 was positive in 34 cases of NPC (34/40, 85%) and 16 cases of nasopharyngeal inflammation (16/20, 80%), without any significant difference between the two kinds of tissues (P>0.05). (2) VIL-10 protein was positive in 30 cases of NPC (30/40, 75%) and 10 cases of nasopharyngeal inflammation (10/20, 50%), with significant differences between the two kinds of tissues (P<0.05). (3) Forty NPC patients were divided into VIL-10 positive and negative groups. The number of CD4 +T cells and ratio of CD4 +T/CD8 + were lower in VIL-10 positive group than in VIL-10 negative group (24.99±3.88% vs. 35.92±6.31%; 0.86 vs. 1.99, P<0.05). The number of CD8 + T cells was greater in VIL-10 positive group than in VIL-10 negative group (30.61±3.34% vs. 20.10±7.68%, P<0.05). Conclusion BCRF-1 gene and its protein product of VIL-10 have high expressions in tissues of NPC. VIL-10 can reduce the number of immune cells in NPC, which will cause immunosuppression. Source


Zhao L.-F.,Head and Neck Tumor Research Center | Li X.-J.,Head and Neck Tumor Research Center | Ma J.,Head and Neck Tumor Research Center | Wang H.,Head and Neck Tumor Research Center | Zhang N.,Head and Neck Tumor Research Center
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2012

Objective To evaluate the effect of short hairpin RNA (shRNA) silencing survivin gene on the apoptosis and proliferation of nasopharyngeal carcinoma CNE-2 cells. Methods The plasmid pSIREN-shRNA/survivin was constructed and transfected into nasopharyngeal carcinoma CNE-2 cells. There were four groups. group A: blank control ones, group B: pSIREN-survivin/nonsense shRNA transfection, group C: pSIREN-shRNA/survivin transfection after 24 h, group D: pSIREN-shRNA/survivin transfection after 48 h. The expression levels of survivin mRNA and protein were detected by RT-PCR and Western blot, respectively. Apoptosis rate and cell cycle were measured by flow cytometry. Cellular proliferation was measured by MTT. Results ShRNA was transfected into CNE-2 cells effectively and transfection rate was (70.9±4.76)% observed with fluorescence microscopy. Survivin mRNA and protein expressions were inhibited in group C, and the inhibition rate was (41.58±0.63)% and (68.29±0.52)%, respectively. The inhibition rate in group D was (63.64±0.96)% and (70.83±0.48)%, respectively. The number of S phase cells decreased while that of G2/M phase cells increased through flow cytometry analysis. The apoptosis rate was (36.24±0.78)% and (50.37±0.85)%, significantly higher than those in the negative and control groups. MTT test showed that the proliferation of CNE-2 cells was inhibited after transfection. Conclusion Short hairpin RNA silencing survivin gene can effectively inhibit expressions of survivin mRNA and protein and induce the apoptosis of CNE-2 cells, thus inhibiting their proliferation. Source

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