Expression Of Sema4D/Plexin-B1 In T Cell Lymphoma And The Biological Behavior

PartⅠThe expression of Sema4 D and Plexin-B1 in patients diagnosed with T-cell lymphomaObjectiveTo investigate the m RNA expression of Sema4 D and Plexin-B1 of T-cell lymphoma paraffin tissue and the solution Sema4 D and Plexin-B1 in serum of T-cell lymphoma.MethodsThe m RNA levels of Sema4 D and Plexin-B1 were detected from paraffin-embedded( FFPE) tissues by real-time quantitative polymerase chain reaction in 21 cases of patients with T cell lymphoma and 10 blocks of Reactive hyperplasia; The Determination of Sema4 D and Plexin-B levels was carried by ELISA from 41 cases’ serum free of T cell lymphoma(24 untreated patients and 17 cases treated patients) and 11 cases of normal control.ResultsThe difference that relative expression level of Sema4 D m RNA(median 3.38, 0.54~11. 35) from T cell lymphoma patients is higher than those reactive hyperplasia(median 0.73, 0.03~1.57), is statistically significant(P= 0.002); There was statistically significant(P=0.028) differences between T cell lymphoma patients(median 3.91, 1.04~37.23) and reactive hyperplasia of Plexin-B1 m RNA relative expression level(median 1.93, 0.58~8.37). The content of serum s Sema4 D from normal persons were 134.51(116.44~218.00) pg/m L, the content of s Sema4 D in serum from the untreated T cell lymphoma patients being 204.20(128.41~259.02) pg/m L, and the content of s Sema4 D in serum was: 157.29(138.52~236.25) pg/m L after treated. The difference between the three groups was statistically significant(P=0.007). The serum s Sema4 D level of untreated T cell lymphoma group is different from the normal person group(P= 0.002); but the serum s Sema4 D level of treated T cell lymphoma group, was the same as the normal control group(P=0.161).The content of serums Plexin-B1 from normal persons were 650.24(416.26~953.58) pg/m L, the content of s Sema4 D in serum from the untreated T cell lymphoma patients being 915.73(499.66~1172.58) pg/m L, and the content of s Sema4 D in serum was 833.07(574.52~1109.45) pg/m L after treated. The difference between the three groups was statistically significant(P=0.046)). Compared with the normal control group, the levels of solution Plexin-B1 in the serum of T Cell Lymphoma Group(untreated or treated) were different(the value of P were 0.044 and 0.022; respectively).ConclusionsThe m RNA relative expression levels of Sema4 D and Plexin-B1 in T cell lymphoma patients were higher than those normal control of reactive hyperplasia; And the levels of solution Sema4 D and Plexin-B1 in serum of patients with T cell lymphoma were higher than those in the control group.PartⅡ The biological behavior regulation of Sema4D/Plexin-B1 signaling pathway in T lymphoma cellObjectiveTo investigate the effect Effects of silencing Sema4 D gene by RNA interference on proliferation, apoptosis and cell cycle of Jurkat cells.MethodsA slow virus Sema4D-RNAi was constructed of by interfering Sema4 D gene. Sema4D-RNAi was transfected into Jurkat cells, and expression changes of Sema4 D m RNA and protein after transfection were detected by real-time quantitative polymerase chain reaction(RT-PCR) and Western blotting. Cell proliferation was analyzed using Cell Counting Kit(CCK-8). Cell cycle and apoptosis of the Jurkat cells were assessed by flow cytometry.ResultsCompared with the normal control group and negative control RNAi group(NC-RNAi group), the relative expression levels of Sema4 D m RNA(1.01±0.01 and 1.01±0.02) were not significantly different(P=0.768). The relative expression levels of Sema4 D m RNA in the Sema4 D si RNA group(A、B and C) after transfection were 0.81±0.06, 0.62±0.03 and 0.62±0.05, respectively, significantly lower than that of the NC-RNAi group(P<0.05 for all). Compared with the normal control group and NC-RNAi group, the expression level of Sema4 D protein(0.99±0.03 and 0.99±0.04) were not significantly different(P=0.907). The expression level of Sema4 D protein in the Sema4 D si RNA group(A, B and C) after transfection were 0.47±0.04, 0.32±0.03 and 0.31±0.02, respectively, significantly lower than that of the NC-RNAi group(P<0.05 for all).The Sema4 D si RNA group C is the best in interferencing, and using this in follow-up study.The proliferation of Sema4 D si RNA group C after transfection was suppressed to some extent, decreased beginning from the 48 hour(P<0.05 for all); and results Compared with NC-RNAi group and normal control group, were not statistically significant(P>0.05). The percentages of G0/G1 cells in the Sema4 D si RNA group C after transfection were(61.43±0.91)%, significantly higher than that of the NC-RNAi group((47.39±0.85)%)(P=0.000). Compared with the normal control group((46.58±0.61)%) and NC-RNAi group, the percentages of G0/G1 cells were not significantly different(P=0.251).The percentages of S cells in the Sema4 D si RNA group C after transfection were(29.38±1.17)%, significantly lower than that of the NC-RNAi group((42.11±1.63)%)(P=0.000). Compared with the normal control group((43.49±1.37)%) and NC-RNAi group, the percentages of S cells were not significantly different(P=0.325).The percentages of G2/M cells in the Sema4 D si RNA group C after transfection were(5.85±0.44)%, the same as the NC-RNAi group((5.30±0.87)%)(P=0.387). Compared with the normal control group((5.18±0.44)%) and NC-RNAi group, the percentages of G2/M cells were not significantly different(P=0.850). The incidence rate of apoptosis in the Sema4 D si RNA group C after transfection were(12.13±0.21)%, significantly higher than that of the NC-RNAi group((2.75±0.16)%)(P=0.000). Compared with the normal control group((2.52±0.15)%) and NC-RNAi group, the incidence rate of apoptosis were not significantly different(P=0.159).ConclusionsDownregulation of sema4 D gene can inhibit the proliferation of human Jurkat cells, promoting apoptosis, and the reason may be arresting cells in mitosis stage G0/G1.