Relationship Between Expression Of MicroRNA-185 And Drug Resistance In Gastric Carcinoma Tissue And Lymph Node Metastases

Objective: Gastric cancer is one of the malignant tumors in human,and the number of newly added gastric cancer in China ranks first in the world.The mortality of gastric cancer is the top three of all cancer mortality,which is highly malignant and postoperative complications seriously reduce the quality of human life.MicroRNA(Micro RNA)is a small molecule that is found in recent years and is related to the gene regulation of non single chain RNA,to participate in the occurrence and progress of a variety of diseases.This topic is based on advanced gastric cancer primary tumor tissue(Primary tumor,PT)and lymph node metastasis(Lymph Node Metastases,LNMs)and the fresh specimens of paraffin block as the research object,using fluorescence quantitative PCR(Real-time PCR RT-PCR)miRNA-185 was detected in PT and LNMs in the expression and the relationship between RT-PCR;technology and immunohistochemical technique(Immunohistochemistry SP)detection of multidrug resistant gastric cancer cell PT and LNMs gene in 1(multidrug resistance 1 gene,MDR1),glutathione S-transferase(glutathione S-transferase-PI GST-,PI),multidrug resistance associated protein-1(multidrug resistance associated protein-1,MRP-1)and thymidylate synthase(thymidylate synthase TS)expression;tetrazolium(MTT)assay techniques commonly used chemotherapeutic drugs(oxaliplatin OXA,Changchun New alkali VCR,5-fluorouracil and paclitaxel)on gastric cancer primary inhibition of tumor cells and metastasis lymph node cells and explore the relation and significance between the tumors and metastatic lymph nodes in mirna-185 expression and tumor drug resistance and drug resistance gene in gastric cancer primary.Methods: The fourth hospital of Hebei Medical University from October 2014 ~2015 October underwent surgical resection for gastric cancer patients with gastric cancer and normal gastric mucosa and lymph node metastasis,a total of 40 patients were confirmed by pathology of lymph node metastasis.Using RT-PCR method for detection of gastric cancer tissues and normal gastric mucosa tissues and lymph node metastasis in mirna-185 expression;using immunohistochemical method for the detection of gastric cancer tissue and metastatic lymph node cells in MDR1 / P-gp,GST,MRP-1 and TS protein expression;gastric cancer chemotherapeutic drugs(oxa,VCR,5-FU,PTX)measurement of gastric primary tumor and metastatic lymph node cells in vitro inhibition by MTT method to detect..all the data were used SPSS 19 statistical software,P < 0.05 that the difference was significant.Results:1 The expression of miRNA-185 in primary gastric cancer,normal gastric mucosa and lymph node metastasis.Expression levels in normal gastric mucosa and gastric carcinoma(0.193 + 154),(0.084 + 0.131),respectively,t=3.752,P < 0.001,the difference have statistical significance;the degree of expression in normal gastric mucosa and metastatic lymph node tissues respectively(0.193 + 154),(0.041 + 0.050),t=6.605,P < 0.001,the difference have statistical significance;the degree of expression in gastric cancer tissues and metastatic lymph nodes were(0.084 + 0.131),(0.041 + 0.050),t=2.129,P < 0.05,both have statistical difference.We can see that the highest expression of miRNA-185 in normal tissues,while the expression in gastric carcinoma and metastatic lymph nodes were decreased.2 Relationship between the expression of miRNA-185 and the expression of multidrug resistance protein(P-gp?MRP-1?GST-??TS)in primary gastric cancer and metastatic lymph nodes.P-gp,MRP-1,GST,TS expression in primary foci of gastric cancer rate was 58.3%(28/48)of the and in 62.5% of 30/48,68.8%(33 / 48),64.6%(more than)in metastatic lymph node positive expression rates were 79.2%(38 / 48),83.3%(40 / 48),75.0%(not),85.4%(48 / 48).Pearson 2 test,P-gp,MRP-1,TS in metastatic lymph nodes in the protein expression was significantly higher than that of the primary tumor(respectively for 2=4.210,P = 0.042;2=4.370,P = 0.038.2=4.140,P = 0.030),GST PI expression strength is higher than that of the primary tumor in the metastases of lymph nodes in the protein,but no significant difference 2=0.840 P=0.483 Bivariate correlation analysis and Pgp,MRP-1,TS protein expression in gastric primary tumor and metastatic lymph node was positively correlated(respectively r=0.448,P = 0.006,r=0.349,P = 0.048,r=0.267,P = 0.031)and GST in primary tumors and metastatic lymph nodes of correlation was found between.In the primary lesions,miRNA-185 and P-gp,MRP-1,TS protein expression was negatively correlated,respectively(r=0.457,P =0.026)(r=0.367,P =0.005)(r=0.248,P =0.034),in the lymph nodes in the various variables were not significantly correlated.3 Comparison of the sensitivity of primary tumor and metastatic lymph node cells to OXA,5-FU,VCR and PTX in vitro chemotherapy.Gastric primary tumor and metastatic lymph node cells,VCR on the metastasis of lymph node cell inhibition rate was significantly lower than that of the primary tumor(t=3.264,P < 0.05),and PTX,5-FU,oxa on tumor cell inhibition rate in gastric primary tumor and metastatic lymph node in no statistical difference(P > 0.05).In the primary tumor cells,the sensitivity of chemotherapy drugs were: PTX > 5-FU > VCR > OXA;the sensitivity of chemotherapy drugs in the metastatic lymph nodes were: PTX > 5-FU > OXA > VCR.4 The relationship between the expression of multidrug resistance protein(P-gp,MRP-1,GST-?,TS)and the inhibition rate of chemotherapy drugs in primary gastric cancer and metastatic lymph nodes.In gastric primary tumor,P-gp strong expression group,5-FU,OXA inhibition of tumor cells was significantly lower than the low expression group(t = 2.467,P <0.05;t = 2.149,P <0.05);MRP-1 strong expression group 5-FU,PTX,VCR inhibition of tumor cells was significantly lower than low expression group(t = 2.553,P <0.05;t = 2.837,P <0.05;t = 2.316,P <0.05);GST-? strong expression of group 5-FU,PTX tumor cell inhibition rate was significantly lower than the low expression group(t = 4.627,P <0.05;t = 3.261,P <0.05);TS strong expression group 5-FU,OXA,PTX on inhibition of tumor cells was significantly lower than if the expression group(t = 5.163,P <0.05;t = 2.479,P <0.05;t = 3.571,P <0.05).In metastatic lymph nodes,P-gp strong expression group PTX,OXA inhibition of tumor cells was significantly lower than the low expression group(t = 5.541,P <0.05;t = 3.168,P <0.05);MRP-1 strong expression group 5-FU,PTX,VCR inhibition of tumor cells was significantly lower than the low expression group(t = 2.261,P <0.05;t = 2.437,P <0.05;t = 681,P <0.05);GST-? strong expression group 5-FU,OXA tumor cell inhibition rate was significantly lower than the low expression group(t = 3.529,P <0.05;t = 3.186,P <0.05);TS strong expression group 5-FU,OXA,VCR tumor the inhibition rate was significantly lower than if the expression group(t = 4.943,P <0.05;t = 2.267,P <0.05;t = 3.142,P <0.05).Conclusion:This study using qRT-PCR technique,cell culture technique,immuno histochemical technique and MTT method on gastric cancer primary mirna-185 expression in tumor and metastatic lymph node cells expression level,resistance protein P-gp,MRP-1,GST,TS protein expression levels and in vitro cell of oxa,5-FU,VCR,PTX drug sensitivity of experiments,and to analyze the correlation.The conclusions are as follows:1 The expression of miRNA-185 in primary gastric cancer and lymph node metastasis was lower than that in normal gastric mucosa,but the expression in metastatic lymph nodes was lower than that in primary tumor.The decrease in the expression level of gastric cancer may be the cause of gastric cancer,and it may be one of the reasons that lead to the increase of related drug resistance protein and decrease the drug sensitivity of tumor cells.2 By immunohistochemical technique to verify Pgp,MRP-1,TS in metastatic lymph nodes in the expression intensity is higher than that of the primary foci of gastric cancer and expression of GST PI in both no statistical difference.3 Gastric primary tumor and metastatic lymph node cells,VCR on the metastasis of lymph node cell inhibition rate was significantly lower than that of the primary tumor(t=3.264,P < 0.05),and PTX,5-FU,oxa on tumor cell inhibition rate in gastric primary tumor and metastatic lymph node in no statistical difference(P > 0.05).In the primary tumor cells,the sensitivity of chemotherapy drugs were: PTX > 5-FU > VCR > OXA;the sensitivity of chemotherapy drugs in the metastatic lymph nodes were: PTX > 5-FU > OXA > VCR.4 P-gp,MRP-1,GST,TS their expression level affect only part of the chemotherapy drug resistance and gastric cancer MDR evaluation should be from the multi-channel,multi factor.In summary,the expression of miRNA-185 in primary tumor and metastatic lymph node was down regulated.The study of drug resistance and drug sensitivity of miRNA-185 may also infer that the expression of multidrug resistance gene may be regulated by the expression of multidrug resistance gene.The results of this study suggest that mirna-185 expression may become a new target for the treatment of gastric cancer,but this study only from the correlation analysis without transfection comparison research method more intuitive,nor the drug resistance related protein mRNA expression level of detection and miRNA-185 expression of MDR specific regulatory mechanism is not entirely clear,pending further in vivo experiments and molecular mechanism research.