Analysis of chromosome aberrations in the cell derived from primary cell culture of laryngeal carcinoma and the Hep-2 cell line / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To search for characteristic chromosome changes in primary laryngeal squamous cell carcinoma (LSCC) and Hep-2 cell line and to realize the relationship between the cytogenetic abnormality and the pathogenetic mechanism in LSCC.</p><p><b>METHODS</b>The fresh resulted samples of LSCC were analyzed with an improved primary cell culture for chromosome preparation and G-banding technique. Hep-2 cell line was analyzed by high resolution banding technique. Molecular cytogenetics analysis was made by chromosome 6 painting probe.</p><p><b>RESULTS</b>Four primary LSCC succeeded in primary cell culture and obtained metaphases, one was tetraploid, the other three were triploid. The chromosome mode of Hep-2 cell line was from 68 to 75 and fifteen marker chromosomes were found. The most structural abnormalities of chromosome in primary LSCC and HEP-2 cell line were unbalance translocation, terminal deletion and isochromosome. The complicate aberration in chromosome 6 was common in LSCC and Hep-2.</p><p><b>CONCLUSION</b>6q-, I(5p), 17p-, 5q- are considered as characteristic chomosome changs in LSCC. Fluorescence in situ hybridization (FISH) may enhance the ability of detecting complicated chromosome rearrangements and marker chromosomes, which could provide more value data to verify the chromosome characteristic aberration in LSCC.</p>

The investigation of STK15 gene amplification and overexpression in laryngeal squamous cell carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the role of STK15 gene amplification and overexpression to genesis and development of laryngeal squamous cell carcinoma (LSCC).</p><p><b>METHODS</b>STK15 gene amplification in 40 cases carcinoma tissues and normal tissues as control was detected by differential PCR approach. STK15 mRNA and protein levels were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry method.</p><p><b>RESULTS</b>In 40 LSCC cases, STK15 gene amplification was found in 14 tumor tissues(35%), mRNA overexpression in 27 tumor tissues(67.5%), and protein upregulated in 29 tumor tissues(72.5%). Statistics analysis showed that STK15 gene amplification and mRNA overexpression were obviously associated to differentiation degree of LSCC, and protein overexpression was closely associated with both differentiation degree and pathological grades of LSCC.</p><p><b>CONCLUSION</b>This research results suggest that STK15 gene amplification contributes to its mRNA and protein overexpression through affecting the exact replication of centrosome and separation of chromosomes. STK15 gene thus plays a role in LSCC oncogenesis and malignant progression.</p>

Mutation of p53 and overexpression of STK15 in laryngeal squamous-cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the relationship between p53 gene mutations and STK15 abnormal expression in the development of human laryngeal squamous-cell carcinoma (LSCC).</p><p><b>METHODS</b>LSCC tissues and matched normal tissues were taken during operation from 55 patients without previous chemotherapy or radiotherapy. Following polymerase chain reaction amplification direct sequencing single strand conformational polymorphism (PCR-SSCP) combined with silver staining were used to detect mutations of p53 gene in exons 7 and 8 (p53E7 and p53E8) using genomic DNA from 110 specimens including 55 LSCC tissues and 55 matched normal tissues. STK15 expression were evaluated by RT-PCR with beta-actin as internal control.</p><p><b>RESULTS</b>The mutation rate of p53E7 was 30.9% (compared to normal tissues, chi(2) = 8.66, P < 0.01). There was no mutation in p53E8. In 38 of the 55 cases (69.1%), the STK15 mRNA expression level was higher than that of the paired normal tissue. The STK15 to beta-actin ratio of average density value was 1.22 +/- 0.49 in the cancer tissue, and 0.99 +/- 0.54 in the normal tissues (t = 4.539, P < 0.01). In 14 of the 17 cases (82.4%) with p53E7 mutations, the STK15 expression was higher than that of normal tissue. In the 38 cases with STK15 over-expression, p53E7 mutation was found in 14 cases (36.8%). The rate of concurrence of p53E gene mutations and STK15 over-expression (25.5%) was higher than that of only p53E gene mutations (chi(2) = 26.025, P < 0.01).</p><p><b>CONCLUSION</b>There is significant association between p53 gene mutation and STK15 over-expression in laryngeal squamous-cell carcinoma.</p>

Deletion of p15 and pl6 genes and overexpression of STK15 gene in primary hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the association of p15 and pl6 genes deletion and STKI5 gene overexpression in primary hepatocellular carcinoma (PHC).</p><p><b>METHODS</b>The carcinoma tissue and the adjacent normal tissue were taken from 30 PHC patients during operations who had had neither chemotherapy nor radiotherapy preoperatively. DNA was extracted from the tissues and PCR was used to determine the homozygous deletion of p15 exon2 (pl5E2) and pl6 exon 2 (pl6E2). RNA was extracted, cDNA was synthesized by RT-PCR, and the expression of STKI5 gene was tested by PCR. Beta-actin was used as an internal control. Average density value (ADV) of STK15 gene and that of beta-actin gene were determined in both carcinoma tissue and the adjacent normal tissue.</p><p><b>RESULTS</b>The rate of p15E2 deletion was 13.3% (4/30) and the rate of p16E2 deletion was 16.7% (5/30) in the carcinoma tissue. The p15E2 and pl6E2 co-deletion rate was 6.7% (2/30). In 19 of the 30 cases (63.3%) the expression of STK15 gene in carcinoma tissue was higher than that in the adjacent normal tissue. The ratio of ADV of STK15 gene to ADV of beta-actin gene (1.53+/-0.31) in the carcinoma tissue was significantly higher than that (0.91+/-0.25) in the paired adjacent normal tissue (t = 2.86).</p><p><b>CONCLUSION</b>The homozygous deletion of p15E2 and p16E2 and overexpression of STKI5 gene may play a role in the oncogenesis and malignant progression of PHC.</p>

Study on STK15 gene abnormality and centrosomal amplification in laryngeal carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate STK15 gene abnormality and centrosomal amplification in laryngeal carcinoma.</p><p><b>METHODS</b>STK15 gene mRNA expressional level was tested in 62 cases of laryngeal squamous cell carcinoma and laryngeal squamous cell carcinoma cell line Hep-2 by reverse transcription-polymerase chain reaction(RT-PCR); the mutation of STK15 gene exon 6 and exon 7 in the same tissues and cells was detected by PCR-single strand conformation polymorphism. Immunofluorescent antibodies were used to test centrosomal amplification in Hep-2 cell line as an example.</p><p><b>RESULTS</b>STK15 gene overexpressed in 39 cases of laryngeal carcinoma (63%) and Hep-2 cell line. No mutation was found in exon 6 and exon 7 of STK15 gene in the above tissues and cells. Centrosomal amplification was apparent in Hep-2 cell line. The number of centrosome in a single cell changed from 1 to 7, and Hep-2 cells with amplified centrosomes (more than 2 in one cell) were 11%-23%.</p><p><b>CONCLUSION</b>STK15 gene overexpression and centrosomal amplification were first found in human laryngeal squamous cell carcinoma, which indicated that STK15 gene overexpression leading to centrosomal amplification might occur in the early stage of human laryngeal carcinogenesis and be one of the key mechanisms for the occurrence of laryngeal carcinoma.</p>