| [Background]Lung cancer is one of the greatest threat in malignant tumors currently, and the incidence appears in rising trend year by year. Use experimental animal models observe the process of tumor development and progression, can shorten research time, explore the etiology and pathogenesis of lun cancer, study the prevention strategies and assess the effect of treatment and so on.[Objective]Explore the soot of Gejiu Tin Smelting Works carcinogenic effects, establish nude mouse model of human lung cancer, then using the model study related protein expression studies, provided experimental evidence to etiology and pathogenesis of Gejiu、Xuanwei lung cancer.[Methods](1) Gejiu refinery smoke vitro induct human bronchial epithelial cells (BEAS-2B) malignant transformation experiments:select high and low concentration group50μg/ml,200μg/ml of YTC refinery smoke exposure immortalized human bronchial epithelial cells(BEAS-2B), each exposed to72h, inter-generational exposure to the9th generation, starting from the10th generation of conventional culture. Take the first30and40generations cells after exposure, inverted phase contrast microscope observe morphological changes in living cells before and after exposure.(2) Lung cancer cell lines in vitro train:recovery Xuanwei lung adenocarcinoma cell line (XLA-07cells), Gejiu lung squamous carcinoma cell line(YTMLC-90cells) and subculture, inverted phase contrast microscope observe the living cells morphology. (3) Nude mice experiment:take high and low concentration group YTC refinery smoke exposure of BEAS-2B first30and40generations cells, human lung cancer cell line XLA-07, YTMLC-90, which were seeded in4-week-old BALB/C subcu-taneously in nude mice, tumor growth was observed in each group situation, measure tumor size and weight of nude mice in every three days, draw tumor growth curve. Kill the mice in the first35days, dissect tissues and organs, the cell morphology was observed with light microscope.(4) Protein expression studies:use immunocytochemistry and immunohistochemistry to test the group of transplanted tumors and cultured cells for Ki-67, MDM2, β-Catenin, Smad2, FHIT, TGF-β1, Smad2.[Results](1) The soot of Gejiu Tin Smelting Works of human bronchial epithelial cells (BEAS-2B), the high concentration200μg/ml which30generations,40generations are able to form xenografts in nude mice, tumor formation rate was100%, compared with non-exposed group and blank control group, the difference was statistically significant (p<0.05); the30th and40th generation in low concentration group50μg/ml which were not tumorigenic, compared with non-exposed and control group, the difference was not statistically significant(p>0.05); high concentration of tumor growth curve was S-shaped; HE staining showed that tissue disorder, many fusiform cells, pathological mitotic common; high concentrations group of the30th and40th generation xenograft FHIT expression was negative, CK17, TGF-β1, MDM2, Smad2expression were positive.(2) The formation rate of Xuanwei lung adenocarcinoma cell line(XLA-07cells) xenograft model in nude mice tumor is60%, the formation rate of Gejiu lung squamous carcinoma cell line (YTMLC-90cells) xenograft model in nude mice tumor is100%, compared with the blank control group, the difference was statistically significant(p<0.05); tumor growth curves of both S-type; HE staining showed that tissue disorder, cell atypia and mitotic figures can be seen more pathological; immunohistochemistry shows that the XLA-07, YTMLC-90xenograft MDM2, β-Catenin were positive, FHIT expression was negative.(3) The Xuanwei lung adenocarcinoma cell line(XLA-07cells) and Gejiu lung squamous carcinoma cell strain (YTMLC-90cells) in immune cell chemical show that the MDM2, β-Catenin expression was positive, the FHIT expression was negative.(4) The immunohistochemistry of Ki67test results:the high concentration YTC refinery smoke exposure immortalized human bronchial epithelial cells positive Sex ratio of30took75percent,40took85percent, and compared with the without smoke exposure of human bronchial epithelial cells, the difference were statistically significant(p<0.05); XLA-07、YTMLC-90cells positive rates were75%,80%respectively. The Xenografts XLA-07、YTMLC-90cells positive rates were83%,89%respectively.[Conclusions](1) The soot of Gejiu Tin Smelting Works could induce malignant of the immorta-lized human bronchial epithelial cells BEAS-2B, the30th and40th generation transformed cells could form transplanted tumor in nude mice.(2) The Gejiu、Xuanwei lung cancer cell lines were implanted into BALB/c nude mice to establish the transplanted tumor model.(3) The proteins of Yunnan Gejiu、Xuanwei regions lung cancer nude mice transplantation tumor, β-Catenin、Smad2、TGF-β1were positive and negative expression of FHIT, The positive rate of Ki67more than70%. |
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