目的 研究人喉表皮癌细胞系Hep-2中乳腺癌易感基因1（BRCAl）、P53结合蛋白1（53BP1）和DNA损伤检测点介质1（MDC1）的表达及临床意义。 方法 采用逆转录聚合酶链式反应检测BRCA1、53BP1、MDC1在喉癌细胞系Hep-2中mRNA的表达，同时用免疫印迹法检测蛋白的表达。 结果 在所检测的人喉癌细胞系Hep-2中BACR1、53BP1、MDC1在基因与蛋白两个水平均有表达。 结论 BRCA1、53BP1、MDC1可能在喉癌的发生发展中有一定作用。
目的 探讨T3期喉癌采用支撑喉镜下CO2激光切除术和部分喉切除术两种手术治疗方式的临床治疗效果。 方法 将2003年8月－2010年7月收治的31例患者按所接受手术方式分为A、B两组（非随机分组），A组16例中男15例，女1例，年龄38～72岁，中位年龄51岁；B组15例，均为男性，年龄46～68岁，中位年龄58岁。病变均累及前联合，A组10例和B组11例累及对侧声带约1/3。A组选择支撑喉镜下CO2激光切除术，B组选择气管切开＋部分喉切除术。两组患者首次术后均未接受放射（放疗）或化学治疗（化疗）。术后第1、3、6、12、24个月门诊纤维喉镜复查。随访时间14～78个月。 结果 A组5例复发或颈部淋巴结转移，复发率31.3%；5例患者均行再次手术、颈清扫及放、化疗。B组4例复发，复发率26.7%；4例均行全喉切除双侧选择性颈清扫术，其中3例术后辅以放、化疗。两组复发率比较差异无统计学意义（P＞0.05）。 结论 采用支撑喉镜下CO2激光切除治疗T3期喉癌，有望得到类似部分喉切除的临床治疗效果。
From Nov. 1988 through Apr. 1994, 78 cases with laryngocarcinoma underwent laryngectomy and laryngeal reconstruction by the use of transfered cervical skin. Three to six months after operation, all patients were subjected to direct or indirect laryngoscope. It was found that the keratinized of layer of the transfered skin tended to be thining out, and that hair had grown in one case. With the followup ranging from 3 months to 5 years,the results were good. All patients following the reconstruction of larynx could phonate, swallow and breath. Postoperatively, 32 patients were alive for 3 years, 2 patients died from recurrence of laryngocarcinoma. The complications included laryngeal fistula in 6 cases and laryngostenosis in 2 cases. The cause as well as the prevention of complications were discussed. It was suggested that thetransfer of cervical flap was feasible for laryngeal reconstruction.
目的:探讨改良气管套管垫安置法对喉癌术后佩戴气管套管、颈部伤口敷料加压包扎期患者的适用性。方法:采用随机分组的方法将38例喉癌术后佩戴气管套管的患者分为传统组20人和改良组18人, 传统组采用“Y”型气管套管垫,改良组采用“Y”型气管套管垫,比较两组患者在更换气管套管垫时的SpO2值、SpO2降低值及刺激性咳嗽次数。结果:安置气管套管垫的过程中,改良组SpO2值高于传统组、SpO2降低值低于传统组,且发生刺激性咳嗽的次数也低于传统组,差异有统计学意义(Plt;0.01)。结论:与传统气管套管垫安置法相比,改良气管套管垫安置法能减少患者换药过程中刺激性咳嗽的次数,对SpO2值影响轻微,更适合于喉癌术后早期颈部伤口敷料加压包扎患者的换药。
目的 构建沉默环氧化酶-2（COX-2）基因重组慢病毒，观察其体外侵袭的抑制作用，从而探讨干扰COX-2抑制喉癌细胞增殖的作用机理，为喉癌的治疗提供新的思路。 方法 逆转录聚合酶链反应（RT-PCR）检测COX-2基因在人表皮样喉癌细胞（Hep-2）中的表达情况。利用上海吉凯公司RNA干扰（RNAi）慢病毒表达载体系统，构建针对COX-2基因慢病毒RNAi表达载体。转染Hep-2细胞，干扰COX-2基因的表达，实时定量PCR检测干扰前后基因表达变化。利用生长曲线测定干扰载体转染前后细胞生长速度变化。流式细胞仪检测细胞的生长周期。Boyden侵袭小室法测定体外侵袭力。 结果 成功构建了COX-2慢病毒RNAi表达载体，并建立了干扰COX-2基因的Hep-2细胞系。实时定量PCR检测COX-2基因在Hep-2细胞系中过表达被显著抑制。生长曲线测定，COX-2基因干扰后细胞增殖明显变慢。流式细胞仪检测细胞的生长周期可见干扰组诱导Hep-2细胞凋亡，转染G0～G1期细胞数量明显上升，S期细胞减少，表明siRNA干扰Hep-2细胞后，细胞由G0～G1期进入到S期受到阻滞，细胞增殖速度下降。体外侵袭实验中，Hep-2-AS侵袭细胞数（31.0 ± 1.8）显著低于Hep-2细胞（104.0 ± 2.6）及Hep-2-P细胞（99.0 ± 2.7），差异有统计学意义（P＜0.05）。 结论 喉癌中过表达的COX-2基因被干扰后表达明显降低并显著抑制细胞的生长速度和侵袭能力。同时验证了COX-2基因RNA干扰在进行抗肿瘤的治疗中潜在的应用前景。
【摘要】 目的 筛选人源喉癌Hep-2细胞株特异结合的短肽，作为喉癌靶向治疗的载体。 方法 体外培养Hep-2细胞株作为靶细胞，人正常喉黏膜上皮细胞为吸附细胞；用噬菌体展示十二肽库进行3轮差减筛选，随机挑取10个噬菌体克隆进行测序；采用酶联免疫吸附（enzyme linked immunosorbent assay，ELISA）法鉴定噬菌体与Hep-2细胞的结合活性；通过免疫荧光鉴定喉癌细胞特异性结合肽（F2）噬菌体阳性克隆与喉癌细胞结合的特异性。 结果 经过3轮筛选后，噬菌体在靶细胞Hep-2上出现明显富集；ELISA分析鉴定显示5个阳性克隆能与Hep-2细胞特异结合，其中F2噬菌体克隆对喉癌细胞的结合靶向性明显高于对照细胞（Plt;0.05）； 免疫荧光显色显示，F2能特异性地与喉癌细胞结合。 结论 利用噬菌体展示肽库技术，可以成功筛选到F2，其可能成为喉癌靶向治疗的载体。【Abstract】 Objective To obtain the polypeptides specifically bound to laryngeal squamous cell carcinoma line (Hep-2) and use it as a potential therapeutic vector targeting laryngeal squamous cell carcinoma patients. Methods With the Hep-2 cells as the target cells and human normal laryngeal squamous epithelial cells (HNLE cells) as the absorber cells, 3 rounds of panning from a Ph.D.-12TM phage-display peptide library were carried out. Ten randomly selected phage clones were sent for sequence detection. The affinity of phage clones was detected by enzyme-linked immunosorbent assay (ELISA). The positive phage clones (F2) specifically bound to Hep-2 were identified by immunofluorescence detection. Results After 3 rounds of screening, 5 positive phage clones showed specific binding to Hep-2 cells and the affinity of positive phage clones (F2) was significantly higher than that of the control groups (Plt;0.05). The results of immunofluorescence detection indicated that F2 could be specifically bound to Hep-2. Conclusions Phage display peptide libraries technique can successfully screen the peptide specifically bound to Hep-2 cell line. Thus, it provides a potential vector for targeting therapy of laryngeal squamous cell carcinoma patients.
Objective To evaluate the expression and clinical significance of Survivin in the tissues of laryngeal carcinoma using meta-analysis. Methods The case-control studies published in China about the expression and association of clinical pathogenic features of Survivin in the tissues of laryngeal carcinoma were electronically retrieved in CBM (1994 to October 2012), CNKI (1994 to October 2012), VIP (1989 to October 2012) and WanFang Data (1996 to October 2012). The reviewers independently identified the literature according to inclusion and exclusion criteria, extracted data, and assessed the quality of the included studies. Then, meta-analysis was performed using RevMan 5.1 software. Results A total of 25 studies were included, involving 1 333 cases of laryngeal carcinoma and 528 cases of health laryngeal mucosa or polyp of vocal cord. The results of meta-analysis showed that, significant differences were found in groups of laryngeal carcinoma vs. health control, laryngeal carcinoma with vs. without lymphatic metastasis, clinical stages I-II vs. III-IV, cell differentiation G1 vs. G2-G3, T1 and T2 stages vs. T3 and T4 stages, and glottic carcinoma vs. non-glottic carcinoma (Plt;0.05). No significant difference was found in groups of age more than 60 vs. no less than 60, male vs. female, and smoke vs. non-smoke (Pgt;0.05). Conclusion Current domestic evidence shows that Survivin may be associated with the whole course of occurrence, advance and transfer of laryngeal carcinoma, and positively correlated to degree of tumor malignance, which may indicate poor prognosis.
ObjectiveTo explore the feasibility to restore pronunciation function by repairing partial suprahyoid epiglottis-preserved circumferential defect in near total laryngectomy with anterior medial thigh flap in advanced laryngeal cancer. Methods A retrospective study of 5 male patients with advanced laryngeal cancer between August 2019 and October 2022, aged 56-73 years, with an average age of 65 years were reviewed. The disease duration ranged from 3 to 24 months, with an average of 8 months. Tumor classification by location: 2 cases of glottic type, 2 cases of supraglottic type, and 1 case of subglottic type; TNM staging: 3 cases of T4N0M0 stage, 1 case of T4N1M0 stage, and 1 case of T4N2M0 stage; American Joint Committee on Cancer (AJCC) staging (2017): stage Ⅳ. Near total laryngectomy with partial suprahyoid epiglottis-preserved and selective bilateral neck dissection were performed before the anterior medial thigh flap was used to repair the circumferential defects. The flap size ranged from 6 cm×5 cm to 8 cm×6 cm. Four patients underwent adjuvant radiotherapy and chemotherapy after operation, while 1 patient did not receive any other adjuvant treatment such as radiochemotherapy. Results The flaps of all 5 patients survived without obvious neck infection. One patient developed a slight pharyngeal fistula after oral feeding at 1 month after operation, which healed after another week of gastric feeding. Primary healing also achieved in the thigh donor area. One patient had bilateral cervical lymph node metastasis, and 1 patient had lymph node metastasis on one side. The remaining 3 patients had no cervical nodes metastasis on both sides. All 5 patients were followed up 12-36 months, with an average of 27.6 months. Four patients had clear, audible, and hoarse voice while 1 patient (case 3) had pronunciation similar to whispering. Laryngoscopy showed that the reconstructed laryngeal inlet was fissure-shape and the reconstructed laryngo-trachea canal below the laryngeal inlet was gradually enlarged. At 1 month after operation, the gastric tube was withdrawn and the food was taken orally. There was no obvious aspiration pneumonia. The tracheostomy tube could be blocked in 4 patients for from 30 seconds to 3 minutes. Among them, 3 patients were able to make a noticeable pronunciation even when the tube was not blocked, and they were able to engage in barrier-free language communication; the tracheostomy tube could not be blocked in 1 patient who had a pronunciation similar to whispering. Preliminary voice analysis showed that the patients have a relaxed and natural pronunciation, without obvious breath-holding or air-swallowing movement, compared to patients with esophageal pronunciation. Decannulation did not achieved until the last follow-up in all 5 patients. ConclusionThe anterior medial thigh flap can repair circumferential defects after near total laryngectomy in advanced laryngeal cancer patients and achieve satisfactory pronunciation, thus can serve as an effective pronunciation rehabilitation method. The preserved part of epiglottis may play a role to prevent postoperative aspiration.