Preterm neonate with a large congenital hemangioma on maxillofacial site causing thrombocytopenia and heart failure: A case report.

BACKGROUND: We report a rare case of a large congenital hemangioma (CH) in the maxillofacial region in a female neonate that caused thrombocytopenia and heart failure. With close multidisciplinary collaboration, the congenital hemangioma was successfully resected with good results. CASE SUMMARY: The patient was delivered at gestational age of 36 wk by cesarean section due to cephalopelvic disproportion and lack of onset of labor (birth weight: 2630 g). A right-sided facial tumor was detected in the fetus during routine antenatal ultrasound examination of the mother at 32 wk of gestation. Physical examination revealed a 7 cm × 7 cm × 3 cm hard, dull purple-colored mass on the right maxillofacial region. The mass was tense and had prominent surface telangiectasias. Laboratory investigations revealed reduced hemoglobin and platelet count, and increased activated partial thromboplastin time, prothrombin time, and thrombin time. International normalized ratio, fibrin degradation products, and D-Dimer levels were significantly increased. Thromboelastography showed increased alpha angle, mean amplitude, and the clot formation speed. Thyroid-stimulating hormone level was significantly elevated. The patient was administered prednisone, propranolol, euthyrox, vitamin K1, milrinone, and digoxin. After operation, cefepime was administered for anti-infection and propranolol was prescribed at discharge. CONCLUSION: We report a rare case of CH in the right maxillofacial region causing thrombocytopenia and heart failure.

Synergistic effect of meta-tetra(hydroxyphenyl)chlorin-based photodynamic therapy followed by cisplatin on malignant Hep-2 cells.

PURPOSE: Tumor drug resistance limits the response to chemotherapy. Interestingly, sequential combination therapy enhances the anticancer efficacy of drugs like cisplatin (CDDP) via synergistic effects. We assayed the synergistic effects of combined photodynamic therapy programmed death receptor-ligand 1 (PDT) and chemotherapy in malignant Hep-2 cells. METHODS: In the cultured Hep-2 cells, meta-tetra(hydroxyphenyl)chlorin (m-THPC) and CDDP were administered separately or in combination. The cellular viability and apoptosis were assessed, accompanied by measurement of the expression of Bax, Bcl-2, ATG-7, and LC3 (LC3-I and LC3-II). Additionally, nuclear chromatin changes, drug retention, and PD-L1 expression were further investigated following different treatments. RESULTS: The sequential treatment significantly diminished cell viability and induced cell apoptosis, in consistency with the usage of single therapeutic strategies, as reflected by an increase in Bax expression and decrease of Bcl-2 expression. Moreover, ATG-7 and LC3-II/LC3-I ratio were reduced after administration of the sequential treatment. Synergetic effect of nuclear chromatin configuration, negative effects of cellular drug retention, and a decrease in PD-L1 expression were observed following the sequential treatment. CONCLUSION: The application of sequential treatment of PDT in combination with chemotherapy offers a promising therapeutic option for cancer treatment, by regulating the PD-L1 expression, autophagy, and non-mitochondrial pathways.

Long non-coding RNA HNF1A-AS is upregulated and promotes cell proliferation and metastasis in nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a common head and neck cancer with an incidence of 10-30 cases per 100,000 in southern China. Although primary treatment includes radiation therapy, prognosis is still unsatisfactory. OBJECTIVE: In this study, we examined the role of HNF1A-AS in NPC progression in vitro and in vivo. METHODS: Relative levels of long non-coding RNA (LncRNA), HNF1A-AS, were evaluated in tumor tissues from 20 patients with NPC as well as from cultured NPC cell lines. Lentivirus-mediated HNF1A-AS knockdown was conducted in NPC cell lines, CNE-2 and SUNE-1. Cell migration and invasion abilities were estimated in vitro by colony-formation, wound-healing, and transwell assays. Cell cycle analysis was used to further examine the role of HNF1A-AS in cell proliferation. The tumor size of 24 male mice with or without HNF1A-AS knockdown was monitored once a week. The underlying mechanism of HNF1A-AS-mediated cell proliferation was studied by western blot analysis. RESULTS: Lentivirus-mediated HNF1A-AS knockdown suppressed cell proliferation and migration abilities. In mice injected with CNE-2 and SUNE-1, depletion of HNF1A-AS caused inhibition of tumor growth, whereas cell cycle analysis showed that HNF1A-AS-knockdown resulted in cell accumulation in the G0/G1 phase. Moreover, HNF1A-AS was found to be associated with epithelial to mesenchymal transition. CONCLUSIONS: Overall, our results suggest that LncRNA, HNF1A-AS potentially regulates NPC tumorigenesis. This could help in development of new strategies for NPC diagnosis and treatment.

Inhibitive effect of IL-24 gene on CD133(+) laryngeal cancer cells.

OBJECTIVE: To explore the inhibitive and apoptosis inductive effect of IL-24 genes on CD133(+) laryngeal cancer cells in Hep-2 line. METHODS: Human peripheral blood monocytes were isolated. The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method. Primers P1 and P2 was designed for the amplification of human IL-24 genes. After confirmation of agarose gel electrophoresis tests, TA was cloned into pMD19-T simple vector. Nhe I and Xho I double digesting human IL-24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector, and detected by enzyme digestion and gene sequencing methods. Flow cytometry (FCM) was used to isolate CD133(+) cells from Hep-2 cells. CD133(+) cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000. After detection, MTT and FCM were used to observe the effect of IL-24 gene on CD133(+) laryngeal cancer Hep-2 cells. RESULTS: Lipotin mediated transfection of recombinant pIRES2-ZsGreen1-hIL-24 plasmid into CD133(+) Hep-2 could expressed IL-24 gene in cells stably. MTT results showed that IL-24 transfected group was significantly suppressed compared to empty vector group and control group (P<0.05); FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group (P<0.05). CONCLUSIONS: IL-24 gene expressions can inhibit proliferation of CD133(+) laryngeal cells in Hep-2 line and promote their apoptosis.

Primary diffuse large B-cell lymphoma of the ethmoid sinus: a case report.

B-cell lymphoma of the paranasal sinuses is rare. We present the case of a 42-year-old woman who presented with proptosis, diplopia, and vision disturbances in the right eye. She was diagnosed with diffuse large B-cell lymphoma of the ethmoid sinus. We describe the general clinical presentation, diagnosis, and differential diagnosis of this entity, and we review the pathology of diffuse large B-cell lymphoma.

Small interfering RNA survivin and GRIM-19 co-expression salmonella plasmid inhibited the growth of laryngeal cancer cells in vitro and in vivo.

OBJECTIVE: To investigate the inhibitory effect of plasmid-based survivin-specific short hairpin RNA and GRIM-19 on the growth of Hep-2 laryngeal cancer cells. METHODS: The plasmid expressing survivin-specific short hairpin RNA (shRNA) and GRIM-19 (p-siRNA survivin/GRIM-19) was prepared and transfected into Hep-2 cells with Lipofectamine 2000. The mRNA and protein expression of surviving and GRIM-19 were measured with RT-PCR and western blot assay, respectively. MTT assay was employed to detect the proliferation of Hep-2 cells, and flow cytometry and AO/EB assay were done to determine the apoptosis of Hep-2 cells. RESULTS: In the p-siRNA survivin/GRIM-19, the mRNA and protein expression of survivin was markedly reduced by 54.4% and 42.2%, and the reduction in protein expression of surviving was more obvious than that in the p-siRNA survivin group (37%) (P<0.05). The protein expression of GRIM-19 was markedly enhanced when compared with the control group (P<0.01). MTT assay revealed the proliferation of Hep-2 cells undergoing transfection with p-siRNA survivin/GRIM-19 was markedly inhibited, and the inhibition rate was as high as 79%, which was higher than that in the psi-survivin group (45%) and p-GRIM-19 group (35%). AO/EB assay and flow cytometry indicated that the apoptotic cells in the p-siRNA survivin/GRIM-19 group were dramatically increased as compared to the psi-survivin group and p-GRIM-19 group. CONCLUSION: The p-siRNA survivin/GRIM-19 has marked decrease in survivin expression and dramatic increase in GRIM-19 expression. Moreover, silencing of survivin and over-expression of GRIM-19 can significantly inhibit the growth and induce the apoptosis of Hep-2 in vitro and in vivo.

Salmonella typhimurium mediated delivery of Apoptin in human laryngeal cancer.

An effective cancer therapeutic should target tumours specifically with limited systemic toxicity. Here, we transformed an attenuated Salmonella typhimurium (S. typhimurium) with an Apoptin expressing plasmid into a human laryngeal carcinoma cell line. The expression of the inserted gene was measured using fluorescence and immunoblotting assays. The attenuated S. typhimurium-mediated Apoptin significantly decreased cytotoxicity and strongly increased cell apoptosis through the activation of caspase-3. The process was mediated by Bax, cytochrome c and caspase-9. A syngeneic nude murine tumour model was used to determine the anti-tumour effects of the recombinant bacteria in vivo. Systemic injection of the recombinant bacteria with and without re-dosing caused significant tumour growth delay and reduced tumour microvessel density, thereby extending host survival. Our findings indicated that the use of recombinant Salmonella typhimurium as an Apoptin expression vector has potential cancer therapeutic benefits.

Chemoresistance of CD133+ cancer stem cells in laryngeal carcinoma.

BACKGROUND: Mounting evidence suggests that tumors are histologically heterogeneous and are maintained by a small population of tumor cells termed cancer stem cells. CD133 has been identified as a candidate marker of cancer stem cells in laryngeal carcinoma. This study aimed to analyze the chemoresistance of CD133(+) cancer stem cells. METHODS: The response of Hep-2 cells to different chemotherapeutic agents was investigated and the expression of CD133 was studied. Fluorescence-activated cell sorting analysis was used to identify CD133, and the CD133(+) subset of cells was separated and analyzed in colony formation assays, cell invasion assays, chemotherapy resistance studies, and analyzed for the expression of the drug resistance gene ABCG2. RESULTS: About 1% - 2% of Hep-2 cells were CD133(+) cells, and the CD133(+) proportion was enriched by chemotherapy. CD133(+) cancer stem cells exhibited higher potential for clonogenicity and invasion, and were more resistant to chemotherapy. This resistance was correlated with higher expression of ABCG2. CONCLUSIONS: This study suggested that CD133(+) cancer stem cells are more resistant to chemotherapy. The expression of ABCG2 could be partially responsible for this. Targeting this small population of CD133(+) cancer stem cells could be a strategy to develop more effective treatments for laryngeal carcinoma.

[Suppressing the growth of Hep-2 human laryngeal cancer cells by silencing survivin gene in vitro and in vivo].

OBJECTIVE: To study the effect of silencing survivin on the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo. METHODS: Hep-2 cells were transfected with pGCsilencer-siRNA-survivin (psi-survivin)by Lipofectamine 2000. The mRNA and protein expressions of survivin were detected by semi-quantitative RT-PCR and Western blot, respectively. Cell proliferation activity was measured by MTT assay. Apoptosis was assessed by flow cytometry. The implanted tumors were formed from injected Hep-2 cells in nude mice. After the tumor formation, psi-survivin was injected into peritumor tissues. The growth of tumor were observed. The tumor volume was calculated and the tumor growth curve was plotted. The expression of survivin in tumor tissues was detected by Western blot. The tumor cell apoptosis was observed by Tunel staining. RESULTS: The sequence-specific siRNA of survivin inhibited the expressions of survivin mRNA and protein. The inhibition rates of survivin mRNA and protein expression were 54.4% and 37.0% respectively. Also the growth of Hep-2 cells was inhibited significantly, with a decrease by 71.7%. By the day 32 of tumor growth, the mean tumor volumes were (1443.9 ± 230.5) mm(3) (x(-) ± s) in saline control group, (1348.5 ± 198.4) mm(3) in plasmid-negative control group, and (624.6 ± 188.4) mm(3) in psi-survivin group, respectively (t = -5.917, P < 0.01). In the implanted tumors injected with psi-survivin, survivin protein expression was down-regulated significantly, with a inhibition rate of 41.8%. Tunel staining showed the apoptosis occurred in the implanted tumors. CONCLUSION: Silencing survivin could significantly inhibit the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo.

[Biological characteristics of highly tumorigenic CD44+CD133+ subpopulation of laryngeal carcinoma cells].

OBJECTIVE: To separate the cell subpopulation with high tumorigenic ability and study the biological characteristics of this subpopulation in laryngeal carcinoma cells. METHODS: Human laryngeal carcinoma cells were obtained by primary tissue culture technique. CD44 and CD133 molecules were used as markers to isolate the CD44(+), CD133(+), CD44(+)CD133(+) and CD44(+)CD133(-) cell subpopulations from the laryngeal carcinoma cells by flow cytometry. A nude mouse tumor xenograft model was developed for the study of the tumorigenic effects of the different cell populations. 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells were injected into the left flank of the mice, respectively. The mice were observed for palpable tumor formation and were sacrificed at 4 weeks later to assess the tumor formation rate, tumor volume and tumor weight. Boyden chamber migration assay was used to determine the migration ability and immunochemistry was used to detect the expression of stem cell antigen SCA-1 and beta1-integrin. Semi-quantities RT-PCR and Western blot analysis were performed to detect the expression level of Bmi-1 in the different cell subpopulations. RESULTS: The growth of subcutaneous tumors in nude mice showed that a tumor can be generated with 1 x 10(3) CD44(+)CD133(+) cells. When the same dose of 1 x 10(6) CD44(+)CD133(+) cells was injected into the mice, both the average weight and volume of the tumors were significantly higher than those generated from other cell subpopulations. Boyden chamber migration assay showed that the invasion ability of CD44(+)CD133(+) cells was significantly higher than that of other cell subsets. The results of immunochemical analysis showed an abundant expression of stem cell antigen SCA-1 and beta1-integrin in the CD44(+)CD133(+) cells. Semi-quantitative RT-PCR and Western blot analysis provided strong evidence that the Bmi-1 expression in CD44(+)CD133(+) and CD133(+) cells was very significantly higher than that in CD44(+), CD44(+)CD133(-) and control cells (P < 0.01). CONCLUSION: Our findings demonstrate that CD44(+)CD133(+) subset cells in laryngeal carcinoma posses some biological characteristics of tumor stem cells, which may be the original cells of laryngeal carcinoma and may become a new target of tumor therapy.

[Preliminary study on biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma].

OBJECTIVE: To study the biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma. METHODS: Tumor samples were obtained from 5 patients, and then the human laryngeal carcinoma cells were cultured in vitro by primary tissue culture technique. Taking CD44 molecule as a marker to isolate CD(44)(+) subpopulation cells from laryngeal carcinoma cells for further study. CD(44)(+) and CD(44)(-) cells were cultured and observed fex their development. CD(44)(+) and CD(44)(-) cells were compared in their functional status (mRNA), cell cycles (G0/G1), their degree of differentiation (CK14 and Involucrin expression) and their morphologic character of the clone. RESULTS: The percentages of CD(44)(+) cells were about 49.8% approximately 53.5% and the median was 51.3%. After culturing CD(44)(+) cells isolated from laryngeal carcinoma could proliferate and the percentage of CD(44)(+) remained the same. CD(44)(+) tumor cells contained much less RNA, more G0/G1 cells, expressed more CK14 protein and less Involucrin protein (less differentiated state). CD(44)(+) cells were multangular in shape with protuberances; CD(44)(-) cells showed a sharp and spindle feature. In comparison with CD(44)(-) cells, CD(44)(+) cells could create heterogeneous offspring by single cell culture of limiting dilution. By observing clone forming rate after single cell planting, it was found that the CD(44)(+) cells had stronger proliferation ability. CONCLUSIONS: CD(44)(+) cells possess some characteristics of stem cells, laryngeal carcinoma stem cells maybe exist in CD(44)(+) cells.

[Construction of the nucleic vaccine pVVP3L-18HN and its antitumor effect on human laryngeal carcinoma].

OBJECTIVE: Nucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes. METHODS: Eukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results. RESULT: The recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours. CONCLUSION: The nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.