Research Of BFGF MAb Combined With Lobaplatin Inhibit The Proliferation And Metastasis And Induce Apoptosis Of Lung Cancer Cells And Its Mechanisms

Objective:(1) To investigate the effect of monoclonal antibodies against b FGF(b FGFm Ab), lobaplatin(LBP), and their combination on proliferation, apoptosis migration and invasion of lung cancer A549 cells, and to provide theoretical support in how anti-b FGFm Ab combined with LBP affect against human lung cancer.(2) To assess the effectiveness and safety of LBP combined chemotherapy for advanced non-small cell lung cancer(NSCLC).Methods: The ascites containing the b FGF m Ab were extracted. The lung cancer A549 cells in logarithm growth phase were selected and then randomly divided into four groups: blank control group, LBP group, anti-b FGF m Ab group, and combination group.(1) CCK-8 assay was used to detect A549 cell proliferation and flow cytometry was used to detect cell cycle in various groups after 48h(of blank control group, LBP group, anti-b FGF m Ab group, and combination group). The expression of protein associated with cell cycle was detected by Western blot.(2) Flow cytometry was used to detect cell cycle in various groups. Nuclear change due to apoptosis was detected with fluorescence microscopy. The expression of protein associated with apoptosis was detected by Western blotting.(3) The function of invasion and migration in A549 was measure by scratch assay and transwell. The expression of protein associated with invasion and migration was detected by Western blotting.(4) Retrieved from Cochrane Library、Pub Med、EMBase、CNKI、CBM VIP database and Wanfang database, RCT about LBP combined chemotherapy versus cisplatin combined chemotherapy for advanced NSCLC were included, the quality of the literature was evaluated by Jadad and valid data was extracted according to inclusionand exclusion criteria, and Rev Man 5.2 software was adopted for Meta-analysis.Results:(1) Cell proliferation was inhibited by LBP and b FGF m Ab, but there was a significant reduction in proliferation in A549 cells treated with a combination of LBP and anti-b FGF m Abs compared with LBP or anti-b FGF m Abs treatment alone(P﹤0.05). Compared with the control group, the cell cycle was arrested at G0/G1 phase and the expression level of CDK4 and cyclin-D was down-regulated.(2) Flow cytometry results showed that b FGF m Abs and LBP could induce the apoptosis of A549 cells and also had synergistic apoptotic effect. A549 nuclear pyknosis, nuclear fragmentation was found by confocal laser scanning microscope after DAPI staining. The expression level of bcl-2, casepase-3, casepase-9, and PARP were down-regulated, where BAX, cleave-casepase-3, cleave-casepase-9, and cleave-PARP were up-regulated.(3) LBP and b FGF m Ab significantly inhibited the migration of A549 cell. The expression level migration associated protein MMP-2 and MMP-9 was down-regulated.(4) A total of 11 RCT were included, involving 724 patients. Meta-analysis showed objective response rate(ORR) [ RR=0.96,95%CI(0.79,1.16), P=0.65], leucopenia[ RR=0.95., 95%CI(0.83, 1.08), P=0.42] and thrombocytopenia [ RR=0.96.,95%CI(0.80,1.15),P=0.68] of two groups had no statistical difference. But trial group could significantly reduced the risks of nausea and vomiting(OR=0.27,95%CI0.18 to 0.45, P<0.05) and diarrhea [ RR=0.43,95%CI(0.31,0.61),P<0.05]than the control group, there was statistical significance.Conclusions:(1)bFGF m Ab combined with LBP inhibit proliferation of A549 cell and the cell cycle was arrested at G0/G1 phase. The mechanism was possibly related with down-regulation of the CDK4 and cyclin-D expression of A549 cell.(2) b FGF m Ab combined with LBP induced the apoptosis and nuclear pyknosis of A549 cell. The mechanism was possibly related with down-regulation of the bcl-2,casepase-3, casepase-9, and PARP expression and up-regulation of the BAX,cleave-casepase-3 cleave-casepase-9, and cleave-PARP by inhibiting the PI3K/AKTpathway.(3) LBP and b FGF m Ab significantly inhibited the migration of A549 cell. The mechanism was possibly related with down-regulation of the protein MMP-2 and MMP-9 expression of A549.(4) Although the effect is similar in the two groups, LBP combination treatment has low side effect than cisplatin combination treatment. Further randomized trials with high-quality and multi center needed because few trials have been included and parts of them are low-quality.