Construction and confirmation of a recombinant adenovirus vector of survivin / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer.</p><p><b>METHODS</b>The survivin gene was cloned by PCR. After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector. The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus.</p><p><b>RESULTS</b>The sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed.</p><p><b>CONCLUSIONS</b>A survivin recombinant adenovirus has been successfully constructed.</p>

Supraglottic horizontal partial laryngectomies in 163 cases / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical value of supraglottic horizontal partial laryngectomy.</p><p><b>METHODS</b>One hundred and sixty-three patients with supraglottic laryngeal carcinoma were treated surgically by supraglottic horizontal partial laryngectomy from 1978 to 1998. There were 64 males and 99 females. Five cases were staged I, 95 staged II, 48 staged III and 15 staged IV. The surgical techniques were improved: The hyoid was removed conventionally; The outer perichondrium of thyroid cartilage was turned into the laryngeal cavity and sutured with the mucosa of laryngeal ventricle and the base of tongue was sutured to the reserved thyroid cartilage. Survival was evaluated using the Kaplan-Meier method. The differences between stages were tested by Los-Rank method.</p><p><b>RESULTS</b>The 5-year survival rate were 100.0%, 77.9%, 54.2% and 33.3% for patients staged I to IV respectively, and were statistically significant (P = 0.0006) between different clinical stages. The 5-year survival rate were 73.1% and 45.5% in patients with cN0 and cN + respectively, and were statistically significant (P = 0.0132). The speech and swallowing functions were good after operation. The decannulation rate was 91.4%. The main causes of death were cervical lymph node metastasis in 40% (20/50) and laryngeal recurrence in 18% (9/50). The occult metastasis rate was 23.1% (30/130) and cervical metastasis rates of patients with cN1-3 75.8% (25/33), with a total metastasis rate was 33.7 (55/163).</p><p><b>CONCLUSIONS</b>Supraglottic horizontal partial laryngectomy is very effective in eradicating disease and in preserving laryngeal function on condition that the indications were selected correctly. The selective lateral neck dissection was recommended for supraglottic carcinoma.</p>

Effect of survivin antisense oligodeoxynucleotide on carcinoma of larynx in vivo and in vitro / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the effect of survivin antisense oligodeoxynucleotide (ASODN) on the apoptosis of human carcinoma of larynx cell line Hep2 and the inhibitory rate in nude mice model so as to discuss the selective blocking activity of antisense technique on gene expression seeking a new way for gene therapy of carcinoma of larynx.</p><p><b>METHODS</b>Antisense oligodeoxynucleotides survivin were transformed into human carcinoma of larynx cell line Hep2 by liposome Lipofectamine 2000. Within 72 h after transfection, 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cellular proliferation. Forty eight hours after transfection, reverse transcription-polymerase chain reaction (RT-PCR) assay was used to observe the expression of survivin gene, Western Blot assay for the protein, and terminal deoxynucleotide mediated nick end labeling (TUNEL) and flow cytometer for cellular apoptosis.</p><p><b>RESULTS</b>Cellular inhibition rate of 72 h went up to 52. 5% and 71.4% at 1.0 micromol/L and 2.0 micromol/L value in Lipo-ASODN groups which differed statistically remarkably (P = 0.046), higher than that in controls in MTT assay (P =0. 003 and 0. 0004). Forty eight hours after transfection survivin gene expression in Lipo-ASODN groups were less than that in control group in RT-PCR assay. Survivin protein expression decreased in Western blot. In TUNEL assay, nuclear positive staining was observed and the apoptosis peak was observed in flow cytometer test, which were absent in controls. In nude mice of carcinoma of larynx model, the inhibitory rate in Lipo-ASODN groups got up to 48.1% and 61.3% higher than that of controls (P < 0.004 and 0. 0006), which differed remarkably (P = 0.032) in a dose-dependently way.</p><p><b>CONCLUSIONS</b>The findings showed that the expression of survivin gene and protein induced cellular apoptosis in Hep2 cells after transfection of Lipo-ASODN and that the carcinoma of larynx in the nude mice model were inhibited by Lipo-ASODN which suggested that antisense technique can be an effective means in the gene therapy of carcinoma of larynx.</p>