Overexpression Of SATB1 In Laryngeal Squamous Cell Carcinoma

ObjectiveTo identify whether SATB1 expressed in laryngeal squamous cell carcinoma and to discuss the clinical significance of SATB1 expression.Materials and Methods1. MaterialsThe study group consisted of 80 patients with LSCC diagnosed and treated between May 2007 and May 2009 at Shengjing Hospital of China Medical University.Among them,25 patients underwent total laryngectomy. Control mucosa samples were obtained from 25 patients who received total laryngectomy, and over 2.0 cm away from the margin of tumor.2. Methods(1) real-time PCRTotal RNA was extracted by Trizol one-step method. After cDNA synthesis, real-time PCR amplication was undertaken.The sequence of the human primers used were as follows:sense:5'-GCA AGC CCT GAG TTC TGT T-3'anti-sense:5'-CTA TGA ATA AGC CTT TGG AGC-3'(2) Western blot analysisSamples were treated with lysis solution to get whole cell protein exracts and then electrophoresed through 10% sodium dodecyl sulfate (SDS)-polyacrylamide gel and were transferred onto a nitrocellulose membrane. Membranes were blocked and incubated with primary antibody. After incubation with alkali phosphatase (AP) conjugated secondary antibody. After treated byDAB, membranes were observed under UVP-8000.(3) ImmunochemistrySP method was used and the procedures were followed by the instruction.(4) Statistical analysisThe data were analyzed by the SPSS software 17.0. Theχ2-test was used to determine the protein expression levels detected by Western blot. Measurement data of relative value of real-time PCR were expressed as mean±SD. Correlations between SATB1 mRNA expression and clinicopathological parameters were also statistically analyzed with T test. Probability values less than 0.05 were considered significant.Results1. All the tumor specimens had SATB1mRNA expression; 64%(16/25) of control mucosa specimens had SATB1 mRNA expression, and other 9 mucosa specimens were absent of SATB1. The difference calculated byχ2 test reached a high level of significance (χ2=27.07; P<0.001). The relative values of SATB1 mRNA in tumor were 5.00±2.48; the values of SATB1 mRNA in control mucosa which had SATB1 mRNA were 0.99±0.88. The mRNA expression levels of SATB1 in LSCCs were 2.52-7.48 fold higher than those in control mucosa tissues; and the difference was statistically significant (P<0.001).2.66.25%(53/80) LSCC specimens had SATB1 protein expression; and none of the mucosa specimens showed detectable SATB1 protein.p-actin expression were similar in all LSCCs and control mucosa samples. The differences were statistically significant (χ2=33.44; P<0.001).3. SATB1 was primarily localized in the nuclei and cytoplasm of the LSCC cells, with no immunoreactivity in control mucosa cells4. The mRNA levels in different sex, age, tumor site, T stage had no statistically difference. The SATB1 mRNA levels in positive cervical lymph node, clinicalⅢandⅣstage, and poor/moderate cell differentiation were significantly higher than those in negative cervical lymph node, clinicalⅡstage and higher degree of cell differentiation.Conclusions1. SATB1 mRNA and protein exist in laryngeal squamous cell carcinomas.2. The mRNA and protein expression levels of SATB1 in LSCCs were higher than those in control mucosa tissues. The difference has statistical significance.3. SATB1 mRNA expression has relationship with clinical stage, degree of cell differentiation and cervical lymph node state.