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Objective:To analyze the expression of LETM2 in KYSE150 and ECA109 cell lines and its effect on the proliferation, migra-tion, and invasion of esophageal squamous cell carcinoma (ESCC). Methods:The expression level of the LETM2 protein in 90 paired hu-man ESCC tissues and matched adjacent normal tissues was determined through immunohistochemistry. The expression level of LETM2 in ESCC cell lines was detected by real-time PCR and Western blot. The expression levels of LETM2 in KYSE150 and ECA109 cell lines were knocked down using lentivirus. MTT assays were performed to examine the effect of LETM2 on the proliferation of ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle. The effect of LETM2 depletion on the migration and invasion of ESCC cells was determined by Transwell assay. Results:LETM2 expres-sion was frequently upregulated in the ESCC tissues than in the adjacent normal tissues. The suppressed exogenous expression of LETM2 led to the inhibition of cell proliferation and colony formation. However, cell migration and invasion were not affected. The re-sults on the cell cycle distribution revealed that LETM2 knockdown acts as a negative regulator of the cell cycle at the G1 to S phase transition. Conclusion:LETM2 acts as a tumor-driven gene in the development and progression of ESCC. This finding suggests that LETM2 can be used as an efficient prognosis biomarker and a potential therapeutic target for ESCC.
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Objective:To analyze the expression of MUC15 and PI3K/Akt in gastric carcinoma and its association with clinicopathologi-cal characteristics and prognosis. Methods:The expression of MUC15 and Akt was detected in 144 cases of gastric carcinoma tissues and corresponding para-carcinoma tissues by tissue microarray and immunohistochemistry. Results:The positive expression rate of MUC15 in gastric carcinoma was 79.8%, higher than that of para-carcinoma tissues (22.2%, P<0.01). The positive expression rate of Akt protein in gastric carcinoma was 80.6%, higher than that of para-carcinoma tissues (16.7%, P<0.01). The expression of MUC15 and Akt was statistically associated with the grades of differentiation, invasion depth, lymphatic metastasis, TNM stage of tumor tissues (P<0.05), and the positive correlation between the two protein expression that appear in the gastric tumor tissue (P=0.001). Univariate survival analysis showed that the over-expression of either MUC15 or Akt was inversely correlated with the survival time (P<0.05 and P<0.01, respectively). Cox multiple regression analysis indicated that patients with over-expression of both MUC15 and Akt had the worst prognoses (HR=3.115, P<0.05). Conclusion:MUC15 may be involved in the occurrence, development, invasion, and metastasis of gastric cancer through the PI3K/Akt cell signaling pathway, and the expression of MUC15 combined with Akt is a powerful predictor for the prognosis of gastric cancer.
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Objective To investigate the expression of Dysadherin and analyze its role in renal clear cell carcinoma (RCCC).Methods RT-PCR and immunohistochemical were used to detect the expression of Dysadherin in 60 cases of fresh RCCC and 60 adjacent normal renal tissues(male 35,female 25; age 37-78,median age 61; ＞7 cm 24,≤7 cm 36; Ⅰ/Ⅱ 39,Ⅲ/Ⅳ 21).Results Dysadherin mRNA expression in RCCC tissues (2.0043±0.2890) was higher than that in adjacent normal renal tissues (0.8461 ±0.2479) (t =6.8020,P ＜ 0.05).Dysadherin expression was associated with nuclear grade.The expression of Dysadherin in nucleus grade Ⅲ and Ⅳ tumors were significantly higher than that in nucleus grade Ⅰ and Ⅱ tumors [the mRNA expression were 4.6224±0.3194,2.7780±0.2288,the positive rates of protein were 64.1％ (25/39),95.2 ％ (20/21) (t =6.5750,x2 =5.495,P ＜ 0.05)].There was no association between the expression of Dysadherin with sex (t =1.0530,x2 =0.023),age(t =0.0511,x2 =0.089) and tumor size (t =1.0330,x2 =0.370) (P ＞ 0.05).Conclusion In RCCC,Dysadherin expression is positively associated with tumor aggressiveness based on grading.It seems that Dysadherin may be a valuable prognostic marker in RCCC.
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The plasmodium of Physarum polycephalum is a suitable eukaryotic cell for cell cycle investigation, but there is no compatible transient expression system for the plasmodium. Using the promoter and terminator of ardC actin of Physarum polycephalum substituted the CMV IE and SV40 polyA of plasmid pDsRedl-N1, using cassette PardC-MCS-DsRed1-TardC substituted the cassette PardC-hph-TardC of plasmid pTB38, we constructed plasmids pXM1 and pXM2 for transient expression of red fluorescent protein (RFP) in Physarum polycephalum respectively. After reconstituting the transcription elongation factor homologous gene (pelf1) of Physarum polycephalum into the pXM2, we generated a plasmid pXM2-pelf1. After the plasmid pXM1, pXM2 and pXM2-pelf1 were electroporated into the plasmodium of Physarum polycephalum, we observed optimum RFP and PELF1-RFP expression under fluoroscope and confocal microscope between 24-48 h after electroporation, and found that ELF1-RFP expression was accumulated in nucleus of microplasmodium, the optimum electroporation parameters were 40 V/cm electric field, 1 ampere current, and 70 micros electric shock time. The results suggest that this expression system is qualified for transient expression of specific protein in plasmodium of Physarum polycephalum.