| Nickel is a kind of metal which is widely contacted in production and life.Occupational exposed to human beings include:nickel of smelting metal purification,plating,making stainless steel and producing battery of nickel and chromium. Accumulated epidemiological evidence and animal experiments confirm that exposure to nickel compounds are associated with tumor, especially lung cancer,the International Agency for Research on Cancer (IARC)classified nickel compounds as group I carcinogen to human in 1990, the crystalline NiS and Ni3S2 are the highest carcinogenic activity among the nickel compounds and the Ni2O3 is second. The carcinogenic actions of nickel compounds are thought to involve four aspects. The first is genomic mutation including ras and p53 induced by nickel compounds, which leads to change the tumor related gene expression pattern and result in carcinogenesis. The second is that nickel induces the signaling pathway in cell leading to transcription factor transactiration by which nickel regulate the expression of specific genes related to tumor development. The third is generation of reactive Oxygen Species (ROS) induced by nickel exposure which leads to DNA and chromatin damage, tumor related gene expression is changed to cause cancer. The fourth, besides nickel induce directly the expression of gene, nickel can change apparent genetics and effect on gene expression of cancer, and at least it will promote cancer development. Recently, numerous epidemiologic and experimental investigations about carcinogenesis and acute lung injury induced by nickel compounds have been reported and some hypotheses have been proposed. However, the exact mechanisms are not known due to its human body. In the present work, the molecular mechanism of lung cancer induced by nickel compounds has been studied on production of ROS, the signaling pathway in cell leading to transcription, inducing NF-κB in cell, intervention effects of Fuzheng Jiedu decoction on molecular mechanism of lung cancer induced by nickel, we try to obtain more understanding on prevention and treatment of occupational lung cancer. 1 Human bronchus epithelial(16HBE)cells in vitro were treated with EME,different concentrations of nickel purite(NiS)(1-8.0μmol/L),NiS(2.0μmol/L),Fuzheng Jiedu Decoction(FJD)(low middle high group),blank serum in 24 hours. We detected survivor rate of 16HBE cells with Trypan—blue tinctured method. 16HBE cells were cultured with culture medium of 24 holes, every hole was 6×106 cells, every dose had five times,16HBE cells were added in all kinds of subjects by different experiment groups and went on culturing 24 hours, group cell was thrown away supernatant fluid, every hole was added in 500μl lytic factor of cells and was put up 30 minutes on the ice,each group cell which was effect on lytic factor was diverted to Eppendorf tube,it was centrifugated 20 minutes,supernatant fluid was detected the expression of glutathione peroxidase (GSH-Px)and reactive oxygen species(ROS). Each group cell which subjects were treated was washed with PBS again and again, until the most cells were broken under microscope, cells were detected activities of superoxide dismutase (SOD).16HBE cells were measured with coomassie brilliant blue,16HBE cells were measured OD of absorbance in 595nm air way with apeotrophotometer and calculated content of protein,16HBE cells were counted content of malondialdehyde (MDA) which was combined with chroma of protein.216HBE cells were inocukated into 25ml culture flask, every hole was inocukated 6×106 cells and was trained for 24 hours,16HBE cells were treated with NiS(1.0μmol/L,2.0μmol/L,4.0μmol/L,5.0μmol/L),MEM of asepsis,NiS(2.0μmol/L)+different FJD serum containing Chinese medicine,blank serum and three kinds of depressant (p38MAPK P-ERK,P-JNK) in 24 hours,16HBE cells were washed two times with PBS and were made in cell suspension after cells were digested with enzyme,16HBE cells were detected the expression of p38MAPK P-ERK,P-JNK with western blot.316HBE cells were treated with different concentration NiS(1.0μmol/L,2.0μ.mol/L,4.0μmol/L),NiS(2.0μmol/L)+different FJD serum containing Chinese medicine in 24 hours,16HBE cells were measured the expression of NF-κB with immunohistochemistry and were analyzed with semiquantitative, 16HBE cells were measured the expression of NF-κB P65 with RT-PCR. This test was analyzed with statistical software of SPSS 13.0,a lot of groups were compared with rariance analysis, two groups were compared with T examination, P<0.05 was regarded as meaning of statistics.Results1 When chroma of NiS was 1.0μmol/L,2.Oμmol/L,4.0μmol/L,.8.0μmol/L, persistence of 16HBE cells were lower than the group of EME, chroma of NiS was higher, persistence of cell was lower, it was 93.6%,92.8%,90.3%,70.3%; the different FJD serum containing Chinese medicine and blank serum were not poisonous to 16HBE cells, persistence of cell was 99.1%,98.8%,99.6%,98.7%.2 NiS can increase obviously the expression of ROS and MDA in 16HBE cells(P< 0.05), chroma of NiS was 1.Oμmol/L,2.0μmol/L,4.0μmol/L, ROS was 31.68U/ml,33.21U/ml,35.76U/ml in 16HBE cells, it existed dose—effect relationship (r=0.8130); the expression of MDA was 0.9518nmol/ml,1.0503nmol/ml,1.1620nmol/ml, it existed dose—effect relationship (r=0.7926), the different FJD serum containing Chinese medicine and NiS (2.0μmol/L)were together treated with 16HBE cells, the expression of ROS and MDA were lower than dyeing nickel group (NiS 2.0μmol/L),middle and high dose FJD were higher than control group(P< 0.05),the blank serum group was not effect on the expression of ROS and MDA in 16HBE cells; NiS inhibited obviously the expression of GSH-Px and SOD (P<0.05), chroma of NiS was 1.0μmol/L,2.0μmol/L,.4.Oμmol/L, the expression of SOD was 11.99 NU/ml,7.89 NU/ml,5.24 NU/ml in 16HBE cells, it existed dose—effect relationship (r=0.8750); the expression of GSH-Px was 22.65 U/ml,15.75 U/ml,10.38 U/ml in 16HBE cells, it existed dose—effect relationship (r=0.8290), the different FJD serum containing Chinese medicine and NiS (2.Oμmol/L)were together treated with 16HBE cells, the expression of GSH-Px and SOD were higher than dyeing nickel group (NiS 2.0μmol/L),middle and high dose FJD were distinguished (P<0.05), the blank serum group was not effect on the expression of GSH-Px and SOD in 16HBE cells.3 NiS can increase obviously the expression of P38MAPK, dose of NiS was higher, the expression of P38MAPK was higher too, the expression of P38MAPK was 0.2868, chroma of NiS was 1.Oμmol/L,2.0μmol/L,4.0μmol/L,5.0μmol/L, the expression of P38MAPK was 0.4112,0.5937,0.6968,0.8976 in dyeing nickel groups.we showed:FJD serum containing Chinese medicine restrained the expression of P38MAPK; middle and high dose FJD were distinguished (P<0.01),but the blank serum group was not any effect on the expression of P38MAPK in 16HBE cells (P>0.05). NiS can increase obviously the expression of P-JNK(P<0.05),the dose of was higher, the expression of P-JNK was higher too,the P-JNK of control group was 0.0410, chroma of NiS was 1.Oμmol/L,2.0μmol/L,4.0μmol/L, the expression of P-JNK was 0.057,0.062,0.068 in dyeing nickel groups, it existed dose—effect relationship (r=0.7736), high dose FJD and SP600125(10μmol/L) restrained the expression of P-JNK(P<0.05), but the blank serum group was not any effect on the expression of P-JNK in 16HBE cells. NiS could not increase obviously the expression of P-ERK1(P>0.05), the expression of P-ERK1 was 0.210 in control group, chroma of NiS was 1.0μmol/L,2.0μmol/L,4.0μmol/L, the expression of P-ERK1 was 0.216,0.219,0.226 in dyeing nickel groups, the different FJD serum containing Chinese medicine and inhibitor of P-ERK1 were not effected on the expression of P-ERK1 (P>0.05).4 NiS could increase obviously the expression of NF-κB and NF-κB P65 (P<0.05),the strong masculine examples of NF-κB in control group were 8, the dose of NiS was 1.Oμmol/L,2.0μmol/L,4.0μmol/L, the strong masculine examples of NF-κB were 17,22,29 in dyeing nickel groups, the middle and high dose FJD serum containing Chinese medicine could descend the expression of NF-κB in 16HBE cells(P<0.05).The optical density of NF-κB P65 in control group was 0.8079, the dose of NiS was 1.Oμmol/L,2.0μmol/L,4.0μmol/L, The optical density of NF-κB P65 in dyeing nickel groups were 0.8558,0.8950,0.9937, the middle and high dose FJD serum containing Chinese medicine could fall the optical density of NF-κB P65(P<0.05). 1 NiS could induce the expression of ROS in 16HBE cells,increase lipid peroxidation and reduce the persistence of cells, the fitting FJD serum containing Chinese medicine could inhibit lipid peroxidation and cut down the expression of ROS in 16HBE cells.2 NiS could induce the expression of P38MAPK and P-JNK in 16HBE cells, but it could not induce the expression of P-ERK1, the fitting FJD serum containing Chinese medicine could reduce the expression of P38MAPK and P-JNK in 16HBE cells.3 NiS could induce the expression of NF-κB and NF-κB P65 in 16HBE cells, the fitting FJD serum containing Chinese medicine could reduce the expression of in 16HBE cells.4 We could show from molecular mechanism of cells:the fitting FJD may be the prevention of the cancer in occupational workers of nickel.
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