Study On M2Pyruvate Kinase (PKM2) As A Target For Molecular Classification In Kazakh’s Esophageal Squamous Cell Carcinoma

Objective: To explore the feasibility of M2pyruvate kinase as Kazakh esophagealsquamous cell carcinoma molecular typing.Methods: At the cellular level, synthetic for PKM2gene siRNA fragments by the Eca109of esophageal squamous cell carcinoma cell lines transfected with Lipofectamine2000.The experiment was divided into three groups, and the blank control group, negativecontrol group, the experimental group. Applied real-time fluorescence quantitativepolymerase chain reaction (real-time quantitative PCR, qRT-PCR) and Western blotanalysis to detect the amount of PKM2expression at the mRNA and protein levels in thecells in each group, and each group was detected by MTT cell viability stream cytometryto detect the changes of the cell cycle and apoptosis, the scratch experiments, and thechanges of Transwell vitro invasion assay migration and invasion.At the tissue level,collected cell carcinoma and adjacent normal tissue which were confirmed examinationKazakh esophageal squamous by surgical resection after pathological examinationconfirmed Kazakh esophageal squamous cell in the First Affiliated Hospital of XinjiangMedical University from2000to2008, using immunohistochemical methods to detectKazakh esophageal squamous cell carcinoma tissue carcinoma and adjacent normal tissuein of PKM2the expression, and analyze the relationship between the analysis andclinicopathological parameters.Results: After the PKM2siRNA successfully transfected Eca109cells, qRT-PCR was usedto detect the three sets of results show that the level of PKM2mRNA expression wassignificantly lower than control group in the experimental group and negative controlgroup, the difference was statistically significant (P=0.012, P=0.018).Western blot analysis showed that PKM2protein expression in the experimental group was reducedcompared with the blank control group and negative group difference was statisticallysignificant (P=0.018, P=0.046). MTT results showed that the proliferative capacity of thecells of the experimental group was significantly weakened, and the other difference wasstatistically significant (P=0.01, P=0.041). After the the PKM2gene expression silencing,apoptosis was66.3%compared with blank control group and negative control group wassignificantly increased, the difference was statistically significant difference (P=0.000,P=0.000). Experimental group cell cycle arrested in G0/G1phase. Transwell invasionassay showed in experimental group the number of cells through the polycarbonatemembrane (56.5±8.7) was significantly lower than the control group (140.3±5.8, P=0.000)and negative control group (109.3±20.1, P=0.036) difference was statistically significant.Scratches experimental results show that cell migration force in the experimental groupwas significantly lower after72hours compared with the other two groups, the differencewas statistically significant (P=0.000, P=0.001). PKM2protein expression in Kazakhesophageal squamous cell carcinoma was significantly higher than the correspondingparaneoplastic tissue, the difference was statistically significant (P=0.043) and hadcorrelated with esophageal squamous cell carcinoma of the depth of invasion (P=0.029),the degree of differentiation (P=0.01), lymph node metastasis (P=0.041) and had nosignificant relationship withwith gender, age, gross type (P>0.05).Conclusions: PKM2can promote proliferation of ESCC cells, accelerate cell invasion,and metastasis.