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KMID : 0857020100250010077
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Kosin Medical Journal
2010 Volume.25 No. 1 p.77 ~ p.89
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Chang in Tumor Growth Factors from Lung Cancer Patient and Nude Mice Injected with Lung Cancer Cell Lines
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Kim Sang-Beum
Park Sung-Dal
Kim Song-Myung
Huh Bang
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Abstract
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Background and purpose : The incidence of non-small cell lung cancer has decreased than past decades but the prevalence rate of squamous cell carcinoma is highest among the various types of lung cancer. There have been one of the most difficult problems of the medical areas because of the law rate of operative treatment by micrometastasis still. The released growth regulating factors from the tumor cells have heen researching targets recently. This study is to know the changes of the tumor growth factors by examining the change of transforming growth factor-β1 (TGF-β1) within plasma of nude mouse was measured after each of different quantity of cultured cell of squamous cell carcinoma was injected into plerual caviry of nude mouse in animal experiment, while comparing the values of TGF, Insulin-like growth factor and
epidermal growth factor between experimental trial and clinical trial.
Method : Between June of 2006 and June of 2007 at Kosin University Hospital, 17 patients were chosen for this study where 13 of them were pathologically diagnosed of squamous cell carcinoma after initial diagnosis of lung cancer followed by radical pneumonectomy and pathologic biopsy, while 4 patients were pathologically diagnosed of benign tumor. Blood sample was collected prior to a surgery from the lung cancer patients scheduled for surgery, and the sample was cryopreserved. During surgery, dense tumor tissue without necrosis was excised in a size of 5mm3 and quickly frozen to be used for experimental tissue, whereas the tissue, which was located farthest from the lesion and deemed free of cancer cell, was to be used for control tissue. And the tissue obtained from benign tumor or normal tissue of granuloma patient was classified as a control tissue B, while the sample from malignant tissue of lung cancer was categorized as experimental tissue M. In animal experiment with 15 male nude mice without immunity, 5 subjects without cancer cell were used for control group, another 5 subjects with of 2×106 of cancer cell line (SW-900 G IV) injected into intrathoracic were used for experimental group I, and the other 5 subjects injected with twice quantity of cancer cell line of group I were classified as experimental group II. After breeding the subjects in the clean room for 8 weeks, blood sample was collected from their heart, and quantitative test of plasma TGF-β was performed. For the test of tumor growth factor, human TGF-β
ELISA kit was used for quantitation of TGF-β1&2, and active TM non-eztraction IGF kit was for quantitation of IGF-I&II, while GE healthcare kit was used for EGF.
Results : In animal experiment, TGF-β1 was expressed within the serum of all nude mice from control group as well experimental group I and II, where control group was at 28.49 fmol/ml while experimental group I and II were at 32.19 fmol/ml and 42.36 fmol/ml respectively. In clinical trial, TGF-β1 was measured higher from experimental group with 40.9 fmol/ml than from control group with 28.5 fmol/ml, and TGF-β2 was lower from experimental group with 12.3 fmol/ml than from control group with 30.3 fmol/ml, while the difference was insignificant with EGF (p＜0.05) where control group and experimental group were at 0.11 ng.ml and 0.12 ng/ml respectively. Moreover, TGF-β1 was at 40.88 fmol/ml from control tissue B and at 15.55 fmol/ml from experimental tissue M, where as TGF-β2 was at 12.31 fmol/ml from control tissue B and 23.95 fmol/ml from experimental tissue M (p＜0.05). TGF-β1 within the serum of control group was at 812.4 fmol/ml before surgery and at 989.1 fmol/ml after surgery, which showed a slight amplification, while TGF-β2 was at 16.8 fmol/ml before surgery and at 20.2 fmol/ml after surgery, exhibiting a slight increase (p＜0.05). Serum IGF-I was at 117.9 ng/ml before surgery and at 214.4 ng/ml after surgery, and IGF-II was at 552.8 ng/ml and 552.8 ng/ml after surgery, while EGF showed insignificant changes between before surgery at 0.67 ng/ml and after surgery at 0.74 ng/ml (p＜0.05). In lung cancer stage Ia,b, TGF-β1 was decreased through surgery as it was at 972.94 mol/ml before and 866.92 mol/ml after, and TGF-β2 was also decreased as measured at 42.40 mol/ml before surgery and 38.24 mol/ml after surgery. In stage IIIa,b, TGF-β1 was decreased as measured at 980.31 mol/ml before surgery and at 928.18 mol/ml, while TGF-β2 showed not much changes (p＜0.05). In lung cancer stage Ia,b, serum IGF-I was decreased as measured at 112.7 mol/ml before surgery and at 87.92 mol/ml after surgery. and IGF-II was also decreased as measured 468.78 ng/ml before surgery and at 358.59 ng/ml after surgery (p＜0.05). In stage IIIa,b, IGF-I showed not much changes, but IGF-II was at 438.31 ng/ml before surgery and 404.2 ng/ml after surgery, where EGF was not all influenced by the phase.
Conclusion : In the experiment with nude mice, TGF-β1 was increased as the number of cancer cell was increased after injection of cancer cell. The number of cancer cell from early tumor, such as Ia,b from post-surgery clinical lung cancer patients, was reduced after the surgery, and TGF-β1 was also reduced as the cancer cell was reduced as well. Therefore, it was believed that TGF-β1 can be a feasible prognostic factor for early diagnosis of lung cancer at Ia,b of early phase.
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KEYWORD
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lung cancer, tumor growth factors, transformed growth factor, insulin-like growth facter, epidermal growth factor
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