The Biological Significance And Expression Of FHIT And Related Protein In Lung Cancer Detected By Tissue Microarray And Cell Microarray

Objective:We studied the expression ofF HIT, MSH2 and p21 protein in progressivelung cancer tissue microarray and cell microarray, and studied theirrelationship with clinicopathological features in priory lung cancerby immunohistocbemical mehtods, furthermore their relationships witheach other were observed. With these results, we hoped to be helpful forgenesis, progress, gene therapy and prognosis evaluation of lung cancer.And to further develop cell microarray and apply a rapid, simple,economical and normative method for the scientific research in cellfields and clinical work.Materials and Methods:The expression of FHIT, MSH2 and p21protein in tissue microarray with270 cores and cell microarray with 112 cores was detected by SPimunohistochemistry method. There were 89 cases of priory lung cancer,12 cases of lymph node metastasis of lung cancer, 12 cases of precancerouslesion and 10 cases of normal lung tissue in tissue microarray and 50cases of lung cancer and 6 cases of norml pleural fluid in cell microarray.All data were processed by SPSS version 11.5 analysis software.Results:1. The expression of FHIT was localized in the cytoplasm. Loss of FHIT expression of primary cancers, precancerous lesion and lymph nodemetastasis of lung cancer.was 46.1%, 41.7% and 50.0% respectively,while 0 in 10 cases of normal tissues. The difference amongfour groupswas significant (p＜0.05). Loss of FHIT expression in precancerouslesion, primary lung cancer and lymph node metastasis of lung cancerwas significantly higher than that in normal lung tissue (p＜0.05).Thedifference among precancerous lesion, primary lung cancer and lymphnode metastasis of lung cancer groups was not statisticallysignificant (p＞0.05). Loss of FHIT expression was related to tumorhistologicol types, degree of cell differentiation and the smokinghistory of patients. (P＜0.05), but not to sex, age, gross appearancetypes, TNM stages, or Lymph node metastasis. (P＞0.05).2. Loss of MSH2 expression of normal tissues, primary cancers,precancerous lesion and lymph node metastasis of lung cancer was 10%, 33.3%, 51.7% and 50.0% respectively. Loss of MSH2 expression inprimary lung cancer was significantly higher than that in normal lungtissue. (p＜0.05) The difference between normal lung tissue andprecancerous lesion, normal lung tissue and lymph node metastasis oflung cancer group, primary lung cancer and lymph node metastasis oflung cancer group, primary lung cancer and precancerous lesion,precancerous lesion and lymph node metastasis was not statisticallysignificant (p＞0.05). Loss of MSH2 expression was related to degreeof cell differentiation tumor and histologicol types (P＜0.05), butnot to sex, age, gross appearance types, TNM stages, or Lymph nodemetastasis. (P＞0.05).3. P21 proteins were respectively detected in 90.0%, 75%, 44% and 25.0% of normal lung tissue, precancerous lesion, primary lung cancer andlymph node metastasis of lung cancer; The positive rate of P21 proteinin normal lung tissue and precancerous lesion were significantlyhigher than that of primary lung cancer and lymph node metastasisof lung cancer (p＜0.05), but the positive rate of P21 protein hadno significant difference between normal lung tissue andprecancerous lesion or between primary lung cancer and lymph nodemetastasis of lung cancer (P＞0.05). In primary lung cancer, theexpression of P21 protein was related to the differentiation degrees,clinical stages and lymph node metastasis. It was higher in the groupof high-middle differentiation than that of low-un differentiation(p＜0.05), and the expression of P21 protein gradually decreased withincreasing malignant degrees of lung cancer; it was higher in thegroup ofⅠ+Ⅱstage than that ofⅢ+Ⅳstage (p＜0.05), and theexpression of P21 protein gradually decreased with the developmentof clinical stages of lung cancer, it was higher in the group withoutlymph node metastasis than that with lymph node metastasis (p＜0.05);But there were no significant difference between the groups ofdifferent age, sex, histological types and gross types (p＞0.05).4. There was significantly positive correlation between FHIT and MSH2(r_s=0.345, p＜0.01), and there was no significantly correlationbetween FHIT and p21 (p＞0.05).5. All points ranks in a good order without distort and distortion incell array. After HE- stained, morphology of the cores was wellpreserved. The diagnosis resulted from the cell array was totally sameto that from the corresponding spread pieces under the microscope. They were answered for the request of research. After IHC stain ofFHIT,MSH2 and p21, the number of cells did not decrease in evidence.The positive expression was totally same to that from the routinespread pieces.Conclusion:1. The protein expression level ofF HIT,msh2 and p21 is reduced duringthe carcinogenesis, infiltrating and metastasis stage of lung cancer,which indicates loss of expression of the three protein mightcorrelate with evolution of lung cancer. The protein expression levelof FHIT is reduced in precancerous tissues, which indicates loss ofFHIT expression might be an early molecular event in lung cancer.The protein expression level of FHIT is reduced in most squamous cellcarcinomas and the patients with a smoking history, which indicatesFHIT could plays a role in the smoking-related carcinogenesis of lungcancer. FHIT expression could be a useful marker for the diagnosisof lung cancer. The expression degrees of MSH2 protein hadsignificantly positive correlation with the differentiation of lungcancer, The lower the differentiated degree is, the higher themalignant degree is. This suggested the loss of MSH2 related withacquired malignant phenotype of tumor cell, it can be areferencein prognosis evaluation. In primary lung cancer, the expressiondegrees of p21 protein had significantly positive correlation withthe differentiation of lung cancer, the expressions of p21 in primarylung cancer gradually reduced following the decrease of tumor celldifferentiated degree. The lower the differentiated degree is, thehigher the malignant degree is. And the expression degrees of p21 protein had significantly positive correlation with clinical stagesand lymph node metastasis, this suggested the loss of p21 relatedwith the development and invasion of lung cancer, it can be a referencein prognosis evaluation.2. There was significantly positive correlation between FHIT and MSH2,loss of their expression promote the genesis and progress of lungcancer, Which suggested there maybe exist cooperation among FHIT andMSH2 protein on the processing of occurrence, progression,invasiveness and metastasis of lung cancer.3. The expression of FHIT, MSH2 and p21 protein in tissue microarray wasdetected by SP immunohistochemistry method, which developed tovalidate that taking 2 cores from the whole-tissue sections was anaccurate and ecomomical method when the diameter of the spot is 0.6mm;and which develop to validate the feasibility and advantage of tissuemicroarray.4. We studied the expression ofF HIT, MSH2 and p21 protein in cell arrayTo further develop cell array. HE-stained and immunohistochemicalstains showing this methodology will be useful for morphology andimmunohistochemical based protein analyses. The cell array canreplace the routine cell spread pieces in the clinic diagnoses andscientific research, especially in the condition of a great many casesneed to deal with.5. Cell array is a new technique, the process is very simple and savingtime, save the materials, reduce the system error. It is rapid, highspecificity, high throughput, so it would be broadly used in theclinical diagnosis and the screen of epidemilology. 6. The cell microarray has its di.sadvantages, such as, it can not providethe formation of histology structure. But They could be used in theresearch about exfoliative cytology and culture cells, and could beused in DNA ploidy analysis et al. Combined to the tissue microarray,it will play more important role in the scientific research.7. Tissue microarray and cell microarray have the advantage of highthroughput, high efficiency, saving reagent and time; it can reducethe experimental error; it has prominent advantage in large scaleresearch...