Cells were transfected with pcDNA expressing CTSL orPLOS A single | plosone.orgOverCaspase 4 Activator site expression of Cathepsin L in Hepatocellular CarcinomaTable 2. Univariate GCN5/PCAF Inhibitor Purity & Documentation survival evaluation of 82 sufferers with HCC.VariablenOverall survival 2-year 5-yearP valueCTSL expression Optimistic Negative Gender Male Female Age(years) ,50 50 Tumor size(cm)D ,five 5 Serum HBsAg Constructive Negative Serum AFP(ng/ml) ,25 25 Cirrhosis Presence Absence UICC stage I+II III+IV Metastasis/Recurrence Yes No Edmondson grade Low (I/II) High (III/IV) D: The largest dimension in the tumor specimen. doi:ten.1371/journal.pone.0112136.t002 62 20 70.three 60.4 43.1 36.8 58 24 50.4 65.two 22.1 34.six 59 23 72.4 54.6 40.1 22.five 64 18 63.4 69.8 32.five 37.2 30 52 72.two 54.1 42.three 21.eight 66 16 62.three 65.1 33.9 37.1 47 35 65.7 52.six 42.1 31.7 58 24 67.six 53.two 48.five 30.3 61 21 62.two 64.1 38.six 40.0 47 35 50.3 64.three 22.7 41.40.0.0.0.0.0.0.0.0.0.Table three. Cox regression evaluation of patients with HCC.VariablesUnivariate HR CI(95 ) two.297,4.067 0.113,1.146 0.412,1.412 0.523,3.736 1.152,3.712 1.218,three.321 0.643,2.789 1.730,six.125 1.121,three.568 0.361,1.P valueCTSL expression(1 = down, 2 = over) Gender(1 = male, two = female) Age(1,50, two 50) Serum HBsAg(1 = damaging, two = positive) Serum AFP(1,25 ng/ml, 2 25 ng/ml) Tumor size(1,five cm, 2 five cm) Cirrhosis(1 = Absence, 2 = Presence) Metastasis/Recurrence(1 = no, two = yes) UICC stage(1 = I+II, two = III+IV) Edmondson grade(1 = High (III/IV), two = Low (I/II)) doi:ten.1371/journal.pone.0112136.t3.025 0.342 0.676 1.466 two.012 1.756 1.365 3.345 2.141 0.0.002 0.091 0.352 0.401 0.017 0.039 0.302 0.000 0.031 0.PLOS A single | plosone.orgOverexpression of Cathepsin L in Hepatocellular CarcinomaFigure 4. The effect of CTSL around the proliferation and tumor progression capability of MHCC-97H cells. A. Western blotting evaluation of CTSL protein expression in HCC cell line (MHCC-97H), colorectal carcinoma cell lines (CaCO2 and LoVo), stably CTSL-expressed MHCC-97H cell line, stably CTSL-expressed CaCO2 cell line, empty vector stable cell lines (MHCC-97H-Con or CaCO2-Con), and MHCC-97H cell line transfected by CTSL-shRNA or Con-shRNA. B. Colony formation assay and MTT assay of MHCC-97H cells and CaCO2 cells with over-expression of CTSL. (Colony formation assay:PLOS One | plosone.orgOverexpression of Cathepsin L in Hepatocellular CarcinomaMHCC-97H-Con (vector) vs MHCC-97H-CTSL, P = 0.0042; CaCO2-Con (vector) vs CaCO2-CTSL, P = 0.0072. MTT: MHCC-97H-Con (vector) vs MHCC-97HCTSL, P = 0.012; CaCO2-Con (vector) vs CaCO2-CTSL, P = 0.035). C. Colony formation assay and MTT assay of MHCC-97H cells with down-regulated CTSL. (Colony formation assay: MHCC-97H-Con-shRNA vs MHCC-97H-CTSL-shRNA, P = 0.003; MTT: MHCC-97H-Con-shRNA vs MHCC-97H-CTSL-shRNA, P = 0.001. (P,0.01 as in comparison to parental groups, P,0.05 as in comparison to parental groups). doi:ten.1371/journal.pone.0112136.gwere sacrificed 21 days after injection. The experiments on mice had been authorized by the ethics committee at Nanfang hospital.Correlation of CTSL Expression with Overall SurvivalThe median follow-up time for general survival was 78 months for all patients. The 2-year and 5-year general prices for all sufferers have been 65.6 and 33.6 , respectively. Among these patients, the overall survival of your sufferers with low CTSL expression (5-year all round price, 41.4 ) was drastically higher than the high CTSL expression group (5-year overall rate, 22.7 ) (P = 0.032, Figure three). Besides CTSL expression level, age, tumor size, serum AFP, s.
