Long noncoding RNA LINC01194 enhances the malignancy of laryngeal squamous cell carcinoma by sponging miR-655 to increase SOX18 expression.

Long noncoding RNAs (lncRNAs) have undergone a comprehensive study for their involvements in tumor treatments. The purpose of our study was to explore the biological effects and regulatory mechanisms of lncRNA LINC01194 (LINC01194) in laryngeal squamous cell carcinoma (LSCC). The levels of LINC01194 in 105 LSCC patients were detected by RT-qPCR. The diagnostic and prognostic value of LINC01194 in LSCC patients were statistically analyzed. The potential functions of LINC01194 in proliferation, apoptosis, and metastasis of LSCC cells were evaluated. The interaction among LINC01194, miR-655 and SOX18 was explored by bioinformatics analysis, luciferase reporter assays and biotinylated RNA pull-down. We found that the expression levels of LINC01194 were highly expressed in LSCC, which was negatively correlated with the clinical outcome of LSCC patients. The area under the ROC curve for LINC01194 was up to 0.8388. Functional assays indicated that LINC01194 knockdown distinctly inhibited LSCC cells proliferation, induced apoptosis, and also attenuated LSCC cells migration and invasion in vitro. Furthermore, we elucidated that LINC01194 promoted SOX18 expression in LSCC cells via functioning as a molecular sponge for miR-655. Overall, based on our findings, LINC01194 served as a tumor promoter and potentially represents a novel prognostic indicator and therapeutic target in LSCC.

TSR2 Induces laryngeal cancer cell apoptosis through inhibiting NF-κB signaling pathway.

OBJECTIVES/HYPOTHESIS: Human laryngeal squamous cell carcinoma (LSCC) is a malignancy that was discovered originally in the epithelial tissue of laryngeal mucosa. However, the underlying molecular mechanism is still not clear. In this study, we aimed to investigate the potential molecular mechanisms of TSR2 in the LSCC cell apoptosis. STUDY DESIGN: The expression of TSR2 was first analyzed in LSCC tissues. Then functional effects of TSR2 on Hep-2 and AMC-HN-8 cell lines were performed by overexpression pcDNA3.1-TSR2. METHODS: We investigated the expression level of TSR2 in LSCC tissues and cells by performing quantitative real-time polymerase chain reaction (qRT-PCR). The pcDNA3.1-TSR2 was constructed to explore the effect of overexpressing TSR2 in Hep-2 cells and AMC-HN-8 cells. We further investigated the effect of overexpressing TSR2 on cell apoptosis-related protein and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 nuclear translocation through Western blot and terminal dUTP nick end-labeling assays. RESULTS: We found that TSR2 was downregulated in LSSC tissues and cells compared with the controls, and the overexpression of TSR2 in Hep-2 and AMC-HN-8 cells could promote cell apoptosis and related apoptosis proteins. The Western blot/qRT-PCR data further indicated that overexpression of TSR2 in Hep-2 and AMC-HN-8 cells could lead to a block of NF-κB signaling pathway via decreasing nuclear NF-κB p65 and increasing cytoplasm NF-κB p65. Moreover, overexpression of TSR2 significantly inhibited the phosphorylation of IκBα and IKKα/ß. CONCLUSIONS: The results indicated that TSR2-induced apoptosis was mediated by inhibiting the NF-κB signaling pathway, which may provide an effective target in gene therapy for LSCC. LEVEL OF EVIDENCE: NA. Laryngoscope, 128:E130-E134, 2018.

Krüppel-like factor 8 is a novel androgen receptor co-activator in human prostate cancer.

AIM: Krüppel-like factor 8 (KLF8) plays important roles in cell cycle and oncogenic transformation. On other hand, androgen receptor (AR) is crucial in development of both androgen-dependent and independent prostatic malignancies. The aim of this study is to investigate the role of KLF8 in prostate cancer (PCa) and the relationship between KLF8 and AR. METHODS: Eight human PCa cell lines, including androgen-dependent LNCap cells and androgen-independent 22Rv1 cells, as well as human PCa samples were studied. LNCap cells and 22Rv1 cells were transfected with plasmids encoding full-length wild-type KLF8 or KLF8 shRNA. The expression of KLF8 protein was detected using Western blotting or immunohistochemical staining. Cell proliferation in vitro was measured with MTT assay, and in vivo in a xenograft nude mouse model. Yeast two-hybrid screening, co-immunoprecipitation and pull down assays were used to examine the binding of KLF8 to AR. Luciferase reporter gene assay was used to measure the transcriptional activity of the genes targeted by AR. RESULTS: In 133 human PCa samples, KLF8 protein staining was observed in 92.65% (63/68) of high-grade PCa, 66.15% (43/65) of low-grade PCa, and 6.82% (3/44) of adjacent normal tissues. The expression of KLF8 was significantly associated with poorer overall survival. Overexpression of KLF8 enhanced the proliferation of both LNCap and 22Rv1 cells, while knockdown of endogenous KLF8 suppressed the proliferation. These manipulations exerted similar effects on the tumor volumes in the xenograft nude mouse model. Yeast two-hybrid screening revealed that KLF8 was a novel AR-interacting protein. With pull down assay and co-immunoprecipitation assay, we demonstrated that KLF8 bound directly to AR, and KLF8 enhanced AR target gene transcription. CONCLUSION: The results demonstrate that KLF8 is a novel AR transcriptional co-activator that is overexpressed in PCa and may play a role in progression of hormone-refractory PCa.

[Construction and confirmation of a recombinant adenovirus vector of survivin].

OBJECTIVE: To construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer. METHODS: 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. RESULTS: 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. CONCLUSIONS: A survivin recombinant adenovirus has been successfully constructed.

[Neck dissection of Levels II and III for the treatment of supraglottic laryngeal cancer with clinically negative neck].

OBJECTIVE: To evaluate the feasibility of routine inclusion of levels II and III in neck dissection to treat the occult neck metastasis as elective treatment for supraglottic cancer with clinically node negative (cN0). METHODS: The results of 52 cN0 patients with supraglottic cancer treated in Tumor Hospital, Harbin Medical University from October 2002 to March 2006 were reviewed retrospectively. RESULTS: Of the 52 patients with supraglottic cancer and cN0 neck, 32 cases received ipsilateral SND (levels II and III) and 20 cases with bilateral SND (levels II and III). Fifteen (28.9%) of 52 patients were found to have occult regional metastasis on pathological examination. Three patients without metastasis in dissected side at pathologic examination showed metastasis in the contralateral undissected neck later on therefore the total occult metastasis rate was 34.6% (18 of 52). The unilateral and bilateral neck occult metastases were determined in 15 cases (28.9%) and 3 cases (5.8%) respectively. A total of 1190 lymph nodes were harvested in 72 specimens, with 30 positive nodes. The distributions of the 30 positive nodes were as follows: level IIA 83.3% (25 nodes), level III 16.7% (5 nodes). Three-year regional recurrence rate was 5.8%. The 3-year survival rate was 84.6% according to Kaplan-Meier in all cN0 patients (n = 52). Patients with positive neck metastasis and patients with extracapsular spread had higher regional recurrence rates (P = 0.021 and 0.002, respectively). CONCLUSIONS: The results support the use of SND (levels II and III) in cN0 supraglottic cancer. This procedure will reduce both operating time and morbidity, without compromising the oncologic result.

[Supraglottic horizontal partial laryngectomies in 163 cases].

OBJECTIVE: To evaluate the clinical value of supraglottic horizontal partial laryngectomy. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

[Cervical metastasis in patients with T2-4 cN0 laryngeal carcinoma].

OBJECTIVE: To study the characteristics of the cervical lymph node metastasis in clinical N0 (cN0) patients with laryngeal carcinoma and its implication in clinical treatment. METHODS: 76 patients with laryngeal carcinomas of T2-4cN0 category were divided into two groups in random: 21(22 sides) radical neck dissection(RND) and 55(60 sides) functional neck dissection(FND) were performed. Lymph nodes were studied histologically according to the levels. RESULTS: On an average, 29.6 lymph nodes were obtained in one side of neck in RND group, and 24.7 in FND group(F = 3.145, P = 0.068). The occult metastasis rates were 33.3% (7/21) in RND group and 34.5% (19/55) in FND group. 25 of 26 patients (96.2%) who had positive nodes involved only the levels II and III. 2130 lymph nodes were obtained in all samples, 59 of 60 positive nodes(98.3%) were located in the level II and III. The 5 and 10-year survival rates of the two groups were 71.4% (15/21), 76.4% (42/55) and 61.9% (13/21), 68.9% (31/45), respectively with no statistical difference(chi 2 = 0.2394, P > 0.5; chi 2 = 0.3143, P > 0.05). Ipsilateral cervical recurrence rates in two groups were 9.5% (2/21) and 7.3% (4/55), respectively with no statistical difference (chi 2 = 0.1059, P > 0.900). 10-year mortalities with negative and positive cervical lymph nodes were 16.7% (7/42) and 62.5% (15/24) respectively, which had statistically difference (chi 2 = 14.4375, P < 0.005). CONCLUSION: The lateral neck (level II, III and IV) dissection may be suitable for the treatment laryngeal carcinoma patients with T2-4cN0.

[Randomized control study on domestic fixed-dose combinations in the initial treatment of smears positive tuberculosis].

OBJECTIVE: To explore the feasibility of domestic fixed-dose combinations as antituberculosis therapy applied in the National Tuberculosis Program. METHODS: A randomized control trial was conducted and 422 smear-positive pulmonary tuberculosis patients were randomly distributed into 2 groups. The trial group was treated daily with rifampicin, isoniazid and pyrazinamide tablets (including 120 mg rifampicin, 80 mg isoniazid and 250 mg pyrazinamide) in the first 2 months and rifampicin and isoniazid tables (including 300 mg rifampicin, 150 mg isoniazid) in the subsequent 4 months. The control group was treated by full 6-month standard regimens (2HRZE/4H3R3). RESULTS: The demographic data and the disease status were similar between the 2 groups before treatment. The rate of conversion from positive to negative cases in the trial group after therapy was 91.6% at 2 months, 97.2% at 3 months, and 97.7% at 6 months, while that of the control group was 87.3%, 97.5%, and 98.0% respectively. The difference in the conversion rates between the two groups was not statistically significant (chi2 = 2. 05, chi2 = 0.03, chi2 = 0.04, P > 0.05). The incidence of skin rash as an adverse effect was less common in the trial group as compared to the control group (Fisher's exact P = 0.024 33) . CONCLUSIONS: Domestic fixed-dose combinations as supplementary anti-tuberculous therapy could be used in the National Tuberculosis Program, but further study is needed for widespread application.