| Objective On the basis of analysis the expression of CD28, CTLA-4, PD-L1andPD-1on T lymphocyte, and expression of Th1, Th2and T lymphocyte relevant factorsIL-12, IFN-γ and IL-10levels, more over detection of tumor regulation genes Bcl-2,NF-κB and TNF-α mRNA levels in Non-small-cell carcinoma (NSCLC) patients, toresearch the mechanism of the changes of T lymphocyte subtypes and relevant controlgenes preoperative and afteroperative of NSCLC patients, to expect provided referencefor clinically prevention, diagnosis and treatment for NSCLC.Methods Blood samples acquired from60NSCLC patients,40patients withbenign lung and50normal control groups, the blood sample of every person weredivided three parts.First part, the first blood samples anticoagulation for detection on percent contentof T lymphocyte and cells level of Th1and Th2by flow cytometer (FCM), and absolutevalue of T lymphocyte by hematology analyzer.Second part, the second obtained serum for detection on T lymphocyte relevantfactors IL-12, IFN-γ and IL-10by enzyme linked immunosorbent assay (ELISA), someof first blood samples anticoagulation for detection on antibodys expression in surfaceof T lymphocyte as CD28, CTLA-4, PD-L1and PD-1and so on.Third part, the third abstracted lymphocyte by lydroxypropylmethyl cellulose fordetection tumor regulation genes Bcl-2, NF-κB and TNF-α mRNA expression levels byReal-time PCR. Results First part, the percent content of CD3+T cells in normal group werehigher than benign group and NSCLC group, in which afteroperation of benign weresignificant lower than preoperation and normal (P＜0.01), NSCLC group andafteropration1-3day group were significant lower than normal group (P＜0.01),NSCLC afteroperation4-7day group were significant higher than benign afteroperationgroup (P＜0.01). The percent content of CD4+T cells in normal group were lower thanbenign group and higher than NSCLC group, in which preoperation and afteroperationof NSCLC were significant lower than normal and benign preoperation (P＜0.01),NSCLC afteropration1-3day group were significant lower than benign afteroperationgroup (P＜0.01). The percent content of CD8+T cells in normal group were lower thanbenign group and NSCLC group, in which preoperation and afteroperation1-3day ofNSCLC were significant higher than normal and benign (P＜0.05)(P＜0.01). Therelative value of CD4+/CD8+in normal group were higher than benign group andNSCLC group, in which preoperation and afteroperation1-3day of NSCLC weresignificant lower than normal and benign (P＜0.01)(P＜0.05).The absolute value of CD3+T cells in normal group were higher than benign groupand NSCLC group, in which afteroperation of benign were significant lower thanpreoperation and normal (P＜0.01), NSCLC group and afteropration1-3day groupwere significant lower than normal group，benign and NSCLC preoperation (P＜0.01),NSCLC afteroperation4-7day group were significant lower than normal group (P＜0.05). The absolute value of CD4+T cells in normal group were higher than benign groupand NSCLC group, in which normal and benign preoperation were significant higherthan benign afteroperation and NSCLC preoperation and after operation (P＜0.01). Theabsolute value of CD8+T cells in normal group were lower than benign group andNSCLC preoperation group and higher than NSCLC afteroperation, in which NSCLCpreoperation were significant higher than normal and benign (P＜0.01), NSCLCafteroperation4-7day were significant lower than preoperation and higher than1-3day(P＜0.05).The absolute value of Th1cells in normal group were lower than benign preoperation and higher than other groups, in which benign preoperation weresignificant higher than normal (P＜0.01), benign afteroperation were significant lowerthan benign preoperation (P＜0.01), NSCLC preoperation and afteroperation weresignificant lower than normal and benign afteroperation (P＜0.01). The absolute valueof Th2cells in normal group were higher than benign preoperation and afteroperationand lower than NSCLC preoperation and afteroperation, in which NSCLC preoperationwere significant higher than normal, benign preoperation and afteroperation (P＜0.05)(P＜0.01), NSCLC afteroperation were significant higher than benign preoperation (P＜0.01). The value of Th1/Th2in normal group were lower than benign preoperationand higher than other groups, in which normal were significant higher than benignpreoperation and higher than NSCLC preoperation (P＜0.01), NSCLC afteroperation1-3day were significant lower than benign preoperation and afteroperation (P＜0.01).Second part, The percent content of CD4+CD28+T cells in normal group were lowerthan benign preoperation group and higher than other groups, in which preoperation andafteroperation1-3day of NSCLC were significant lower than normal and benignpreoperation (P＜0.01). The percent content of CD8+CD28+T cells in normal group werehigher than other groups, in which normal were significant higher than preoperation andafteroperation of NSCLC (P＜0.01). The percent content of CD8+CTLA-4+T cells innormal group were lower than other groups, in which preoperation of NSCLC weresignificant higher than normal (P＜0.01), afteroperation4-7day of NSCLC weresignificant lower than preoperation (P＜0.01). The percent content of CD4+PD-L1+Tcells in normal group were higher than benign afteroperation and lower than othergroups, in which preoperation of NSCLC were significant higher than normal (P＜0.05).The percent content of CD8+PD-L1+T cells in normal group were lower than othergroups, in which benigen preoperation and NSCLC preoperation and afteroperation4-7day were significant higher than normal (P＜0.05)(P＜0.01), NSCLC preoperationwere significant higher than benigen preoperation (P＜0.01). The percent content ofCD4+PD-1+T cells in normal group were higher than NSCL afteroperation4-7day andlower than other groups, in which NSCLC preoperation and afteroperation1-3day were significant higher than normal (P＜0.05)(P＜0.01), NSCLC afteroperation4-7daywere significant lower than preoperation and afteroperation1-3day(P＜0.01)(P＜0.05).The percent content of CD8+PD-1+T cells in normal group were lower than other groups,in which NSCLC preoperation and afteroperation1-3day were significant higher thannormal (P＜0.01)(P＜0.05).The absolute value of CD4+CD28+T cells in normal group were lower than benignpreoperation group and higher than other groups, in which benign afteroperation andNSCLC preoperation and afteroperation were significant lower than normal (P＜0.01)(P＜0.05), NSCLC afteroperation1-3day were significant lower than preoperation(P＜0.05), NSCLC afteroperation4-7day were significant higher than1-3day(P＜0.05).The absolute value of CD8+CD28+T cells in normal group were higher than other groups,in which normal were significant higher than preoperation and afteroperation of NSCLC(P＜0.05)(P＜0.01). The absolute value of CD8+CTLA-4+T cells in normal group werelower than other groups, in which preoperation of NSCLC and afteroperation1-3daywere significant higher than normal (P＜0.01). The absolute value of CD4+PD-L1+Tcells in normal group were lower than other groups, in which preoperation of NSCLCafteroperation1-3day were significant higher than normal (P＜0.05). The absolutevalue of CD8+PD-L1+T cells in normal group were lower than other groups, in whichNSCLC preoperation and afteroperation1-3day were significant higher than normal (P＜0.01). The absolute value of CD4+PD-1+T cells in normal group were lower than othergroups, in which NSCLC preoperation and afteroperation were significant higher thannormal (P＜0.01)(P＜0.05). The absolute value of CD8+PD-1+T cells in normal groupwere lower than other groups, in which NSCLC preoperation and afteroperation weresignificant higher than normal (P＜0.01), NSCLC preoperation were significant higherthan benign afteroperation (P＜0.01).the standard curve of IL-12ELISA was very well. The value of IL-12in normalgroup were lower than benign preoperation and higher than other groups, in whichbenign preoperation were significant higher than normal (P＜0.05), NSCLCpreoperation and afteroperation were significant lower than normal, benign preoperation and afteroperation (P＜0.05)(P＜0.01), NSCLC afteroperation1-3day were significantlower than preoperation (P＜0.05), NSCLC afteroperation4-7day were significanthigher than afteroperation1-3day (P＜0.05).The standard curve of IFN-γ ELISA was very well. The value of IFN-γ in normalgroup were higher than benign afteroperation and lower than other groups, in whichNSCL preoperation and afteroperation4-7day were significant higher than normal,benign preoperation and afteroperation (P＜0.05), NSCLC afteroperation1-3day weresignificant lower than preoperation (P＜0.05).The standard curve of IL-10ELISA was very well. The value of IL-10in normalgroup were higher than benign preoperation and afteroperation and lower than othergroups, in which benign preoperation were significant lower than normal(P＜0.05),NSCLC preoperation and afteroperation4-7day were significant higher than normal,benign preoperation and afteroperation (P＜0.05)(P＜0.01), NSCLC afteroperation1-3day were significant higher than benign preoperation and afteroperation and NSCLCpreoperation (P＜0.05).Third part, the amplification curve and melting curve of Bcl-2, the amplificationefficiency and specificity were very well. The level of Bcl-2gene in normal group werelower than benign and NSCLC preoperation and afteroperation, in which normal weresignificant lower than other groups (P＜0.01), NSCLC preoperation were significanthigher than benign preoperation and afteroperation (P＜0.01), NSCLC afteroperationwere significant lower than NSCLC preoperation (P＜0.01).The amplification curve and melting curve of NF-κB, the amplification efficiencyand specificity were very well. The level of NF-κB gene in normal group were lowerthan benign and NSCLC preoperation and afteroperation, in which normal weresignificant lower than benign preoperation and NSCLC (P＜0.01), NSCLCpreoperation were significant higher than benign preoperation and afteroperation (P＜0.01), NSCLC afteroperation were significant lower than NSCLC preoperation (P＜0.01).The amplification curve and melting curve of TNF-α, the amplification efficiency and specificity were very well. The level of TNF-α gene in normal group were lowerthan benign and NSCLC preoperation and afteroperation, in which normal weresignificant lower than NSCLC (P＜0.01), NSCLC preoperation were significant higherthan normal, benign preoperation and afteroperation (P＜0.01), NSCLC afteroperation4-7day were significant higher than benign preoperation and afteroperation, NSCLCpreoperation (P＜0.01).Conclusion1The immune system about T lymphocyte were abnormally inNSCLC patients, and improved afteroperation. The percent content of CD3+T andCD4+T were down regulation, and the percent content of CD8+T were up regulation, andCD4+/CD8+were down regulation in patients.2The immunity was down regulation in patients, the CD8+CD28+T cells and Tccells were decrease. The relevant co-stimulatory molecule of CTLA-4, PD-L1and PD-1were up regulation at different degree.3The ratio of Th1and Th2was disbalance in NSCLC patients, in which the benigntoward Th1and the NSCLC toward Th2.4In the body of NSCLC patients the immuno-regulation factors could be upregulation for the cancer cells, although immune escape mechanism was in cancer cells,and the IFN-γ and IL-10were up regulation but IL-12.5The immune system and cancer cells both carry the game in NSCLC patients, inwhich TNF-α high levels. The cancer cells influence apoptosis gene expressions, theanti-apoptosis gene Bcl-2high expression so as NF-κB, it could be profit for cancercells live in bodys. |
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