e Knockdown of TM4SF1 inhibited SKOV3 cells migration. xenograft tumors in nude mice were examined. Results (1) The positive expression rate of TM4SF1 protein in epithelial ovarian cancer tissues (90.90%) was higher than that in benign ovarian tumor tissues (65.22%) and normal ovarian epithelial tissues (31.25%), and both differences were significant (benign ovarian tumor tissues, normal ovarian epithelial tissues Primers and short hairpin RNA (shRNA) Primers were designed using the Primer 5 primer design software according to the coding sequence (CDS) of human TM4SF1 (“type”:”entrez-nucleotide”,”attrs”:”text”:”NM_014220″,”term_id”:”1519242232″,”term_text”:”NM_014220″NM_014220) in GenBank. Primers sequences are presented in Table?2. Table 2 Primers sequences valuevalue
FIGO3.8680.012?I~II2116 (76.2)?III~IV3434 (100. 0)Histological grade4.4620.035?Grade 11612 (75.0)?Grade 2C33938 (97.4)Histological typeC0.398?Serous2927 (93.1)?Mucosity1615 (93.8)?Endometrioid86 (6/8)?Clear cell22 (2/2)Ascites0.0190.892?<500?ml3733 (89.2)???500?ml1817 (94.4) Open in a separate windowpane Statistic by Fishers exact probability test The association between the positive expression rate of TM4SF1 protein in epithelial ovarian malignancy cells and clinical prognosisUnivariate and multivariate analysis results showed the FIGO stage and histological grade were both influencing factors of ovarian malignancy patient prognosis, and positive TM4SF1 protein expression was not an independent element affecting the prognosis of ovarian malignancy individuals (P?>?0.05) (Table?6). Table 6 Univariate and multivariate analyses of factors influencing prognosis of ovarian malignancy
Age0.81 (0.32C1.64)0.368CCFIGO2.92 (1.53C6.06)0.0031.60 (0.97C9.40)0.032Histological grade1.89 (0.93C3.57)0.0100.96 (0.61C6.34)0.042Ascites1.07 (0.58C2.29)0.0461.02 (0.79C4.05)1.193TM4SF12.02 (1.00C7.89)0.0031.12 (0.68C9.46)1.047 Open in a separate window The expression of TM4SF1 in HO8910PM and SKOV3 cells after RNAi Testing of siRNA fragments that experienced the best silencing effect using RT-qPCRDifferent gene silencing efficiencies were recognized using fluorescence quantitative PCR. The results showed that all 3 of the siRNA constructs experienced inhibitory effects (P?Rabbit Polyclonal to NOX1 gene and protein in HO8910 after RNAi. c, e Manifestation of TM4SF1 gene and protein in SKOV3 cells after RNAi. *: p?P?P?Fendiline hydrochloride S phase was 32.16??3.01, and the percentage of cells in G2 Fendiline hydrochloride phase was 14.61??4.32; these ideals were not significantly different from those of the bad control group (LV-CON-RNAi-Luc/HO8910PM) (Fig.?4a, b) . Open in a separate windowpane Fig. 4 The effect of RNAi on cell cycle of HO8910PM and SKOV3 cells. a Cell cycle of LV-CON-RNAi-Luc/HO8910PM cells. b Cell Fendiline hydrochloride cycle of LV-TM4SF1-RNAi-Luc/HO8910PM. c Cell cycle of LV-CON-RNAi-Luc/SKOV3 cells. d Fendiline hydrochloride Cell cycle of LV-TM4SF1-RNAi-Luc/SKOV3 cells The circulation cytometry results showed the percentage of cells in G1 phase in the LV-TM4SF1-RNAi-Luc/SKOV3 group was 30.74??1.82, the percentage of cells in S phase was 42.77??0.66, and the percentage of cells in G2 phase was 26.49??1.48; these ideals were not significantly different from those of the LV-CON-RNAi-Luc/ SKOV3 group (Fig. ?(Fig.4c,4c, d). The effect of RNAi.