The Expressions Of MRP-1 MRNA And Protein In Lung Cancer Progression Tissue Microarray And Research Of Corresponding Molecule Mechanism

Objective: To investigate the role of MRP-1/CD9 in occurrence, development, invasiveness and metastasis of lung cancer and corresponding molecule modulating mechanism, we detected its mRNA and protein level and related marker KAI1.FAK and NF-κB protein expressions in a lung cancer progression tissue microarray and analyzed the relationship between they and clinical parameters. We hope to supply a theoretical evidence for diagnosis, genetic therapy and judgment of prognosis.Material and methods: We applied Fluorescence in situ hybridization technique and SP immunohistochemical method to detect mRNA and protein level of MRP-1/CD9 and KAI1, FAK and NF- κB protein expressions and analyzed their relationship each other in a lung cancer progression tissue microarray containing normal lung tissue specimen, precancerous diseases, cancer specimen and metastasis specimen by. All data were processed by SPSS version 11.0 analysis software. Results:1. The result of factured TMA: The character of our recipient block was exquisite and had no splits. The chip cores were arranged orderly and clearly in the block except one core had shift. The chip cores had no shift and distort and obviously rugate in slides when they cutted from block. They were answered for the request of research.2. The mRNA expression level of MRP-1 was gradually reduced following tumor progression. The positive rate was 100.0% in 10 cases of normal tissues and58.3% in precancerous tissues, 38.2% in primary cancers and 8.3% in metastasis cancers, the difference among four groups was significant (p<0.01) Thereinto the differences between normal tissues and premalignant lesions and primary cancers respective were significant (P<0. 01) , the difference between premalignant lesions primary cancers and was opposite (P>0. 05) .3. The protein expression level of MRP-1 was gradually reduced following tumor progression. The positive rate was 100.0% in 10 cases of normal tissues and 58.3% in precancerous tissues, 41.6% in primary cancers and 8.3% in metastasis cancers, the difference among four groups was significant (p<0.01) Thereinto the differences between normal tissues and premalignant lesions and primary cancers respective were significant (P<0. 01) , the difference between premalignant lesions and primary cancers was opposite (P>0. 05) . The mRNA expression level of MRP-1 was consistency with protein. The difference was insignificant (P>0.05) .4. The protein expression of MRP-1 in every histological type group was different. Squamous cell cancer>adenocarcinoma>large cell cancer, there was no expression in SCLC group. The difference among four groups was insignificant(pX).O5) . Combined into NSCLC and SCLC group, the difference was significant ( p<0.01 ) The MRP-1 protein expression was different in respectively differentiated group. The level was gradually reduced following the decrease of differentiated degree. The expression gradually reduced following tumor clinical progression. The expression level in the group without lymph node metastasis was higher than control group. All the differences were significant (p<0.05) . Furthermore, The expression of MRP-1 had no relationship with other parameters.5. The expression level of KAI1 was gradually reduced following tumor progression. The positive rate was 100.0% in 10 cases of normal tissues and 66.7% in precancerous tissues, 24.7% in primary cancers and 0.0% in metastasiscancers. The difference between normal tissues and premalignant lesions was insignificant (P>0. 05), the differences between normal tissues and primary cancers, premalignant lesions and primary cancers were opposite (P<0. 05) . combined normal and precancerous as non-neoplasm group, the difference among three groups was significant (p<0.01)6. The expression of KAI1 in primary lung cancers had no relationship with age and gender of the patients and the macroscopic type of tumor (p>0.05) . The protein expression level of KAI1 in every histological type group was different. Squamous cell cancer> large cell cancer > adenocarcinoma> SCLC. The difference among four groups was insignificant (pX).O5). But in NSCLC group the difference among three groups was significant (p<0.05) . The KAI1 protein expression was different in respectively differentiated group (well, moderate, low. undifferentiated). The expression level in high-moderate group was higher than low-undifferentiated group (p<0.05) . The expression gradually reduced following tumor clinical progression. There was significance difference among them(p<0.01). The expression level in the group without lymph node metastasis was higher than control group. The difference was significant (p<0.01) .7. The expression level of NF- k b was gradually increased following tumor progression. The positive rate was 0.0% in 10 cases of normal tissues and 33.3% in precancerous tissues, 36.0% in primary cancers and 66.7% in metastasis cancers. The difference among four groups was insignificant (p,X).O5) . Thereinto the difference between normal tissues and premalignant lesions was significant(P<0. 05) , the difference between normal tissues and primary cancers was significant , too (P<0.05) . The difference between premalignant lesions and primary cancers was opposite (P>0. 05) .8. The expression of NF- k B in primary lung cancers had no relationship with ageand gender of the patients and the macroscopic type and histological type of tumor (p>0.05) . The expression of NF- k b gradually increased following tumordifferentiated degree decreased. There was significance difference among them (P<0. 05) . The NF- k B protein expression was different in respectively clinicalstage group. The difference was significant (P<0. 05) . The expression hadrelationship with tumor whether accompany lymph node metastasis (P<0. 01) .9. The expression level of FAK was gradually increased following tumor progression. The positive rate was 10.0% in 10 cases of normal tissues and 16.7% in precancerous tissues, 48.3% in primary cancers and 83.3% in metastasis cancers, the difference among four groups was significant (p<0.05). Thereinto the difference between normal tissues and prem'alignant lesions was insignificant(P>0.05) , the differences between normal tissues and primary cancers, premalignant lesions and primary cancers were opposite (P<0. 01) .10. The expression of FAK had no relationship with other parameters such as age, gender, macroscopic type (p>0.05) . The protein expression level of FAK in every histological type group was different, adenocarcinoma > SCLC > squamous cell cancer > Large cell cancer. The difference among four groups was insignificant(pX).O5) . The expression level of FAK was related with differentiated degree in primary lung cancer. FAK expression in high-moderate group was lower than that in low-undifferentiated group, the difference was significant (p<0.01) . The expression gradually raised following tumor clinical progression. Furthermore, The positive rate in the group with lymph node metastasis was higher than control group (p<0.01) .11. There was significantly positive association between KAI1 expression and MRP-1 expression (rs=0.521, P<0.01). there was significantly negative response between KAI1 and FAK (rs=-0.256, P<0.05) . There was no relationship betweenKAI1 and NF- k B (rs=0.195, PX).O5). MRP-1 was negatively associated with FAK and wasn't associated with NF- k B (rs==0.401, PP<0.01; rs==0.200, PX).O5). FAK wasn't associated with NF-k b ,too (rs==0.075, PX).O5) . Conclusion:1. The mRNA level and protein level of MRP-1 in premalignant lesions and primary cancers were both decreased. This indicated it participate in the occurrence of tumor. Its abnormal expression may be early event. Detecting the marker will help to forecast occurrence of lung cancer. The loss of MRP-1 in metastasis lesions may explain its consanguineous relationship with metastasis function. The expressions of MRP-1 in different histological types of lung cancer were various. The difference of KAI1 expression between SCLC and NSCLC was no significant. However, expression of MRP-1 was loss in SCLC, compared with that in NSCLC, the difference was significant. These suggested MRP-1 may play important role in SCLC target therapy.2. The expressions of MRP-1 in primary lung cancer gradually reduced following the decrease of tumor cell differentiated degree. The lower the differentiated degree is, the higher the malignant degree is. This suggested the loss of MRP-1 related with acquired malignant phenotype of tumor cell.3. The expression of MRP-1 in primary lung cancer was related with clinical stage, which indicated it participate in the malignant progression of lung cancer. It's obviously down expression implied lung cancer late stage. Detecting the marker may supply important information for estimate patient's prognosis. The expressions of MRP-1 in tumors with lymph node metastasis were lower than those without lymph node metastasis in primary cancer group. This indicated MRP-1 play important role in course of inhibiting cancerous cell invasion and metastasis and the loss of MRP-1 related with acquired malignant phenotype of tumor cell.4. The expressions of KAI1 and MRP-1 in primary lung cancer were lower than that in normal and precancerous groups and higher than that in metastasis group. The expression level of the two proteins kept high consistency. Furthermore there was positive response. These suggested KAI1 and MRP-1 participate in the occurrence, progression, invasion and metastasis of lung cancer and they cooperated with each other.5. The expressions of KAI1 in primary lung cancer gradually reduced following the decrease of tumor cell differentiated degree and the progression of clinical stage. This suggested KAI1 participate in the differentiation and malignant progression of tumor cell. The expressions of KAHin tumors with lymph node metastasis were lower than those without lymph node metastasis in primary cancer group. This indicated KAI1 play important role in course of inhibiting cancerous cell invasion and metastasis.6. There was no expression of NF- k B in normal lung tissue The positive rate in precancerous diseases was 33.3% and 36.0% in lung caner and 66.7% in metastasis tissues. The expression level was increased in precancerous group, which indicated NF- k b participated in occurrence of cancer. Its abnormal was early event. The expression level related with differentiated and clinical stage. Detected the marker will important to estimate cancer progression. There was no significant relationship between NF- k b and other three marker respectively. The mechanism on NF- k B regulate lung cancer need to study again.7.The expression of FAK in primary lung cancer was higher than that in normal and precancerous groups and lower than that In metastasis group, which suggested FAK protein participate in the processing of occurrence, progression, invasiveness and metastasis of lung cancer and play an important role. There was negative response respectively between FAK and KAI1, FAK and MRP-1, Which suggested...