Predictors of quality of life in primary caregivers of patients with heart failure: A model of health literacy and caregiving burden.

BACKGROUND: Heart Failure (HF) is a chronic disease that impairs patients' ability to care for themselves. The accumulation of caregiving activities by caregivers to patients creates stress. OBJECTIVES: This study intends to investigate the mediating role of caregiving burden in the relationship between health literacy and quality of life of caregivers. METHODS: This study is a cross-sectional research conducted through a questionnaire survey. A convenience sampling method was employed to select 410 primary caregivers for the study. RESULTS: The overall mean score for quality of life for caregivers of patients with HF was (49.30±9.64). The results showed that the caregiving burden mediated the relationship between health literacy and quality of life, with the mediating effect accounting for 39.04 % (P < 0.05) of the total effect. CONCLUSION: Caregiving burden is a mediating variable in the relationship between health literacy and quality of life. Therefore, we offer some recommendations for healthcare professionals: ①We suggest that healthcare professionals provide relevant education and training to caregivers, as this can enhance their knowledge and skills in effectively managing the health condition of patients;②Healthcare professionals can also proactively assess the caregiver's burden level and design personalized support plans based on the assessment results.

Constructing nursing quality indicators for intraoperative acquired pressure injury in cancer patients based on guidelines.

Cancer patients have a high incidence of intraoperative acquired pressure injury (IAPI). Constructing IAPI quality indicators can reduce the incidence of pressure injury, but there are a lack of these indicators targeting cancer patients. Based on this, this study develops a system of quality indicators for IAPI. Thirty-four potential indicators were included based on the literature review. The 26 experts were asked to rate the importance and feasibility of each indicator using three rounds of email survey. The authoritative coefficient ranged from 0.92 to 0.94. After three rounds of Delphi expert consultation, nine nursing quality indicators were identified for IAPI in cancer patients. The mean importance or feasibility ratings ranged from 4.77 to 5.81 on a six-point scale, with variation coefficients ranging from 0.07 to 0.26. The percentage of full score for potential indicators ranged from 23.10% to 80.80%. Over three rounds, the Kendall's W coefficients ranged from 0.157 to 0.354 (P < .01). The absolute and relative importance and feasibility of the nine indicators were identified as potentially valid measures of nursing quality indicators for IAPI in cancer patients. This instrument is the first set of IAPI quality indicators developed specifically for cancer patients, and it should be useful for evaluating and improving the quality of IAPI in this population.

Correlation of signal transducer and activator of transcription-3 and ß-catenin expression in laryngeal squamous cell carcinoma.

OBJECTIVE: The specific roles of phosphorylated signal transducer and activator of transcription-3 (p-STAT3) and ß-Catenin in laryngeal squamous cell carcinoma (LSCC) remain unclear. METHODS: In this study, the correlations between p-STAT3, ß-Catenin, and clinicopathological characteristics were investigated using tissues and clinical data from 124 LSCC cases. Immunohistochemistry and immunofluorescence assays were used to examine p-STAT3 and ß-Catenin expression and localization in these samples. Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognostic significance of these proteins. LSCC cell lines were treated with a STAT3 inhibitor (dihydroartemisinin) or activator (interleukin-6) to explore the mechanism of p-STAT3 and ß-Catenin. RESULTS: There was an inverse correlation between p-STAT3 and ß-Catenin expression in the LSCC samples. Patients with high p-STAT3 and low ß-Catenin expression levels had significantly worse overall survival. Multivariate Cox regression analysis revealed that lymph node metastasis and ß-Catenin expression were both independently correlated with unfavorable overall survival. Cell treatment with the p-STAT3 inhibitor inhibited the nuclear accumulation of ß-Catenin, while p-STAT3 activator treatment could promote ß-Catenin translocation to the nucleus. CONCLUSION: Overall, our data indicate that p-STAT3 expression is associated with LSCC by promoting ß-Catenin degradation.

Dihydroartemisinin inhibits IL-6-induced epithelial-mesenchymal transition in laryngeal squamous cell carcinoma via the miR-130b-3p/STAT3/ß-catenin signaling pathway.

OBJECTIVE: To explore whether dihydroartemisinin (DHA) can block interleukin (IL)-6-induced epithelial-mesenchymal transition (EMT) in laryngeal squamous cell carcinoma (LSCC). METHODS: The expression of SLUG, signal transducer and activator of transcription 3 (STAT3), and microRNA (miR)-130b-3p was measured. In addition, a dual-luciferase reporter assay was performed to examine the interaction of miR-130b-3p with STAT3. RESULTS: We found that IL-6 can promote EMT and invasion in LSCC cells, whereas DHA can inhibit these two processes. However, DHA alone does not influence EMT and cancer invasion. Furthermore, DHA upregulated miR-130b-3p, which can downregulate STAT3 and ß-catenin protein expression and decrease the activity of the IL-6/STAT3 signaling pathway. Moreover, we found that miR-130b-3p can target STAT3 directly. CONCLUSIONS: DHA can block IL-6-triggered EMT and invasion in LSCC, and during these processes, DHA increases miR-130b-3p expression to decrease the activation of the IL-6/STAT3 and ß-catenin signaling pathways. These findings may provide new insights into strategies for suppressing and even preventing LSCC metastasis.

Correlation analysis of sacrococcygeal pressure and operation time in patients undergoing general anesthesia in the supine position.

OBJECTIVE: This prospective study was performed to explore the change in sacrococcygeal pressure during an operation under general anesthesia in the supine position and identify the correlation between pressure injury and body mass index. METHODS: This study involved 99 patients who underwent general anesthesia. Sacrococcygeal pressure was measured and recorded at seven time points: before general anesthesia, 5 minutes after general anesthesia, and 1, 2, 3, 4, and 5 hours after the beginning of the operation. The pressure change at each time point was compared, and the factors affecting the pressure were analyzed. RESULTS: The correlation analysis showed that the operation time was significantly and positively associated with the occurrence of pressure injury. CONCLUSION: Perioperative management should be strengthened to speed up the surgical process and shorten the operation time, which will help to reduce the occurrence of intraoperative pressure injury.

Application of the Caprini risk assessment model for deep vein thrombosis among patients undergoing laparoscopic surgery for colorectal cancer.

ABSTRACT: Application of the Caprini risk assessment model was explored in patients with deep vein thrombosis (DVT) after laparoscopic colorectal cancer surgery.This study was a prospective study. The risk factors for DVT were assessed with a survey at baseline and on the morning of surgery, first day after surgery and sixth day by using repeated blood vessels on color Doppler ultrasound of the lower limbs, and the intraoperative and postoperative conditions were recorded.Among 148 surgical patients, 24.3% had asymptomatic DVT. According to the risk stratification, the incidence of DVT was related to the Caprini score (P < .001). The area under the curve of the Caprini model was 0.701 ±â€Š0.047 (95% CI: 0.609-0.793, P<.001). The Youden index was 0.368, while the critical point was 10.5 in the Caprini model, corresponding to a sensitivity of 0.806 and a specificity of 0.563. Age, cardiovascular disease, intraoperative blood loss, postoperative fever, preoperative preparation, and hospital stay were higher in DVT patients than in patients without DVT. Moreover, the incidence of DVT in patients with a lithotomy position was higher than that in patients with a scissors position. In binary logistic regression analysis, the independent risk factors for DVT development were age, intraoperative blood loss, and preoperative preparation time.The Caprini model can be used for the prediction of venous thromboembolism in laparoscopic colorectal cancer surgery patients. The thrombosis risk assessment model must be established in line with patients undergoing endoscopic malignant tumor surgery.

Dihydroartemisinin Prevents Progression and Metastasis of Head and Neck Squamous Cell Carcinoma by Inhibiting Polarization of Macrophages in Tumor Microenvironment.

BACKGROUND: Polarized M2 macrophages are an important type of tumor-associated macrophage (TAM), with roles in the growth, invasion, and migration of cancer cells in the tumor microenvironment. Dihydroartemisinin (DHA), a traditional Chinese medicine extract, has been shown to inhibit the progression and metastasis of head and neck squamous cell carcinoma (HNSCC); however, the effect of DHA on cancer prevention, and the associated mechanism, has not been investigated in the tumor microenvironment. MATERIALS AND METHODS: First, human Thp-1 monocytes were induced and differentiated into M2 macrophages using phorbol 12-myristate 13-acetate (PMA), interleukin-6 (IL-6), and interleukin-4 (IL-4). Induction success was confirmed by cell morphology evaluation, flow cytometry, and quantitative real-time polymerase chain reaction (qRT-PCR). Then, DHA was applied to interfere with M2 macrophage polarization, and conditioned medium (CM), including conditioned medium from M2 macrophages (M2-CM) and conditioned medium from M2 macrophages with DHA (M2-DHA-CM), was obtained. CM was applied to Fadu or Cal-27 cells, and its effects on cancer invasion, migration, and angiogenesis were evaluated using transwell, wound-healing, and tube formation assays, respectively. Finally, Western blotting was used to evaluate the relationship between signal transducer and activator of transcription 3 (STAT3) signaling pathway activation and M2 macrophage polarization. RESULTS: Human Thp-1 monocytes were successfully polarized into M2-like TAMs using PMA, IL-6, and IL-4. We found that M2-like TAMs promoted the invasion, migration, and angiogenesis of HNSCC cells; however, DHA significantly inhibited IL-4/IL-6-induced M2 macrophage polarization. Additionally, as DHA induced a decrease in the number of M2-like TAMs, M2-DHA-CM inhibited the induction of invasion, migration, and angiogenesis of Fadu and Cal-27 cells. Finally, DHA inhibited M2 macrophage polarization by blocking STAT3 pathway activation in macrophages. CONCLUSION: DHA inhibits the invasion, migration, and angiogenesis of HNSCC by preventing M2 macrophage polarization via blocking STAT3 phosphorylation.

Dihydroartemisinin Prevents Distant Metastasis of Laryngeal Carcinoma by Inactivating STAT3 in Cancer Stem Cells.

BACKGROUND Accumulating evidence indicates that cancer stem cells (CSCs) are a minor subpopulation of cancer cells that may be the primary source of cancer invasion, migration, and widespread metastasis. MATERIAL AND METHODS We investigated the effects of dihydroartemisinin (DHA) on distant metastasis of laryngeal carcinoma and the relevant mechanism. In vitro, we used the Hep-2 human laryngeal squamous carcinoma cell line (Hep-2 cells) to assemble CSCs, using CD133 as the cell surface marker. Our data demonstrate that the CD133⁺ subpopulation of Hep-2 cells has greater invasion and migration capabilities than CD133⁻ cells. We also evaluated the effects of DHA, a newly defined STAT3 inhibitor, on the invasion and migration of CD133⁺ Hep-2 cells under hypoxia and IL-6 stimulation, both of which can activate STAT3 phosphorylation. RESULTS CSCs exhibited a significant decrease in the ability of migration and invasion upon the application of DHA, along with simultaneous alterations in related proteins, both in cultured cells and in xenograft tumors. The associated signaling proteins included phosphorylated STAT3 (p-STAT3), matrix metalloproteinase-9 (MMP-9), and E-cadherin, which are closely involved in cancer invasion and metastasis. In vivo, we found that DHA can reduce lung metastasis formation caused by CSCs and prolong survival in mice, and can inhibit STAT3 activation, downregulate MMP-9, and upregulate E-cadherin in lung metastatic tumors. CONCLUSIONS Taken together, our findings indicate that CSCs possess stronger invasive and metastatic capabilities than non-CSCs, and DHA inhibits invasion and prevents metastasis induced by CSCs by inhibiting STAT3 activation.

Heavy Metal Contamination and Ecological Risk Assessment of Swine Manure Irrigated Vegetable Soils in Jiangxi Province, China.

Heavy metal are often added to animal fodder and accumulate in the soils with swine manure. In this study, heavy metal (Cu, Pb, Cd, Zn, As and Cr) concentrations were determined in agricultural soils irrigated with swine manure in Jiangxi Province, China. Results showed that the average concentrations of Cu, Zn, As and Cr (32.8, 93.7, 21.3 and 75.8 mg/kg, respectively) were higher than the background values, while Pb and Cd (15.2 and 0.090 mg/kg, respectively) were lower than the background values. Contamination factors [Formula: see text] indicated that they were generally moderate for Cu, Zn, As and Cr and generally low for Pb and Cd. The contamination degree (C d ) was calculated to be 7.5-10.0 indicating a moderate degree of contamination. The geoaccumulation index (Igeo) indicated that the soils were unpolluted with Zn, Cd and Pb, while unpolluted to moderately pollute with Cr, Cu and As. The single ecological risk factor [Formula: see text] revealed that the six heavy metals all belonged to low ecological risk. The ecological risk indices suggested that all the sampling sites were at low risk level.

[Combined inhibition of STAT3 and HIF-1α for enhancement of chemosensitivity in the model of human laryngeal squamous cacinoma in nude mice].

OBJECTIVE: To investigate the effects of combined inhibition of signal transducer and activator of transcription 3 (STAT3) and hypoxia-inducible factor-1α (HIF-1α) in the enhancement of chemosensitivity of the model of human laryngeal squamous cacinoma in nude mice. METHOD: Model nude mice were divided into six groups randomly: control group(A) , cisplatin group(B) , cisplatin and AG490 group(C) , cisplatin and HIF-1α⁻/⁻ group (D), cisplatin combined AG490 and HIF-1α⁻/⁻ group (E), HIF-1α⁻/⁻ group (F) (only in calculating tumor inhibition rate). 3mg/kg cisplatin was administered by peritoneal injection for 3 days. Then cisplatin and 10 mg/kg AG490 were administered every other day for 12 days. The expression of Ki67 and HIF-1α was detected by immunocytochemical method. Western blot was used to detect the expression of p-STAT3. RESULT: The expression of HIF-1α in group C and group D were lower than that in group B, and there were significant difference respectively (t1 = 2.782, t2 = 3.873, P < 0.05); The expression of HIF-1α in group E was lower than that in group C and group D respectively, and there were significant difference respectively (t1 = 6.140, t2 = 3.667, P < 0.01). The expression level of p-STAT3 in group C was markedly lower compared with that in group B, and there were significant difference between them (t = 17.840, P < 0.01); There were no difference between the expression level of p-STAT3 in group D and that in group B (t = 0.038, P > 0.05); The expression level of p-STAT3 in group E was significantly lower compared with that in group C and group D respectively (P < 0.01). Tumor inhibition rate of group E was higher than that in group B, group C , as well as group D respectively and there were significant difference respectively (t1 = 5.509, P < 0.01; t2 = 3.422, P < 0.05; t3 = 2.718, P < 0.05 ). Ki67 index of group E was lower than that in group B, group C as well as group D respectively and there were significant difference respectively(t1 = 8.307, P < 0.01; t2 = 3.736, P < 0.05; t3 = 4.524, P < 0.01). CONCLUSION: Combined inhibition of STAT3 and HIF-1α could enhance chemo-sensitivity in the model of human laryngeal squamous cacinoma in nude mice.

Study on the effect and mechanism of the dysfunction of CD4(+) T cells in the disease process of chronic cardiac failure.

OBJECTIVE: To study the effect and mechanism of the dysfunction of CD4(+) T cells in the disease process of chronic cardiac failure (CHF). METHODS: According to different group technologies, 100 CHF patients were divided into the following groups: ischemia group and non-ischemia group, heart function Ⅲ-Ⅳ group and heart function Ⅰ-Ⅱ group, event group and non-event group, and 50 healthy volunteers were included in the control group. Real-time PCR was used to detect transcription factors T-bet and GATA-3 of Th1 and Th2; flow cytometry was applied to determine the ratio of Th17 and Treg cells; ELISA was employed to test cytokines IFN-γ, IL-4, IL-17 and IL-10 of peripheral blood Th1, Th2, Th17 and Treg cells, respectively; ultrasonic cardiogram was used to exploit to LVEF and LVEDd; and electrochemilu minescene immunoassay was used to examine plasma BNP. The differences of all indexes of all groups were analyzed and the correlation between CD4 T cells and clinical indexes was analyzed by Pearson correlation analysis. RESULTS: As compared to the control group, the transcription factors T-bet and GATA-3 of Th1 and Th2, the ratio of cytokines Th17 and IFN-γ, cytokines IL-17, T-bet/GATA-3, IFN-γ/IL-4, Th17 cells/Treg cells, IL-17/IL-10 of the ischemia group and non-ischemia group, heart function Ⅲ-Ⅳ group and heart function Ⅰ-Ⅱ group, event group and non-event group were all increased significantly, while their transcription factor GATA-3 of Th2, cytokines IL-4, Treg cells ratio, cytokines IL-10 were decreased obviously. The differences showed statistical significance (P < 0.05). The increase or decrease of the partial CD4+ T cells of the ischemia group, heart function Ⅲ-Ⅳ group and event group was more distinctly. The results of Pearson correlation analysis showed that IFN-γ and IL-17 were significantly positively correlated with LVEDd and BNP, IL-4 and IL-10 were also significantly positively correlated with LVEF, but correlated negatively with BNP, and IL-17 was negatively correlative with LVEF. CONCLUSIONS: There was a correlation between CHF and the dysfunction of CD4(+) T cells showing immune activation phenomenons of deviations from the Th1/Th2 balance towards Th1 and from the Th17/Treg balance towards Th17, which was also related to the types, severity and prognosis of the disease.

Dihydroartemisinin as a Putative STAT3 Inhibitor, Suppresses the Growth of Head and Neck Squamous Cell Carcinoma by Targeting Jak2/STAT3 Signaling.

Developing drugs that can effectively block STAT3 activation may serve as one of the most promising strategy for cancer treatment. Currently, there is no putative STAT3 inhibitor that can be safely and effectively used in clinic. In the present study, we investigated the potential of dihydroartemisinin (DHA) as a putative STAT3 inhibitor and its antitumor activities in head and neck squamous cell carcinoma (HNSCC). The inhibitory effects of DHA on STAT3 activation along with its underlying mechanisms were studied in HNSCC cells. The antitumor effects of DHA against HNSCC cells were explored both in vitro and in vivo. An investigation on cooperative effects of DHA with cisplatin in killing HNSCC cells was also implemented. DHA exhibited remarkable and specific inhibitory effects on STAT3 activation via selectively blocking Jak2/STAT3 signaling. Besides, DHA significantly inhibited HNSCC growth both in vitro and in vivo possibly through induction of apoptosis and attenuation of cell migration. DHA also synergized with cisplatin in tumor inhibition in HNSCC cells. Our findings demonstrate that DHA is a putative STAT3 inhibitor that may represent a new and effective drug for cancer treatment and therapeutic sensitization in HNSCC patients.

[Combined inhibition of Stat3 and HIF-1α for enhancement of radio- and chemo-sensitivity in laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the influence of signal transducer and activator of transcription 3 (Stat3) and hypoxia-inducible factor-lα (HIF-1α) on the resistance effect of laryngeal squamous cell carcinoma to radiation therapy and chemotherapy under the hypoxia circumstances. METHOD: Western blot was used to test the expression of p-Stat3 and HIF-1α in the Hep-2 cells under the hypoxia conditions. MTT assay was used to test the proliferation of Hep-2 cells after radiation therapy and chemotherapy; the Hep-2 cells were suppressed expression of Stat3 and/or HIF-1α. RESULT: (1) AG490 induced significant proliferation inhibition on Hep-2 cells and Hep-2HIF-1α-/- cells in vitro underthe hypoxia environments (P < 0.05); (2) Suppressing expression of Stat3 reduced the expression of HIF-1α protein (P < 0.05); (3) Combined inhibition of Stat3 and HIF-1α enhanced radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypoxia. CONCLUSION: Combined inhibition of Stat3 and HIF-1α can further enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypixia compare than inhibiting Stat3 or HIF-1α alone. Effectively blocking of HIF-1α pathway and suppressing the expression of Stat3, would be an effective method to enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells, which provides a new thought to reduce the resistance to treatment.

Interaction between TNFR1 and TNFR2 dominates the clinicopathologic features of human hypopharyneal carcinoma.

Although the expression of tumor necrosis factor receptors (TNFRs) has been associated with clinicopathologic features of some other cancers, their roles in hypopharyngeal squamous cell carcinoma (HPSCC) have not been documented. Forty-five HPSCC specimens were analyzed for the expression of TNFR1 and TNFR2 and its relationship with clinicopathologic factors. Interaction between the two receptors and its effects on TNF-α was investigated by neutralizing TNFR1 and upregulation of TNFR2. The results indicated that, in HPSCC specimens, the expression of TNFR1 but not TNFR2 is associated with clinical staging, T stage, cervical lymph node metastasis, and histologic grade in HPSCC. In Fadu cells, when conjugating with its receptors, TNF-α mediates proliferation effects, and neutralizing TNFR1 and/or upregulating TNFR2 evokes proliferation-inhibiting and apoptosis-inducing effects and potentiates cisplatin (DDP)-induced growth inhibition and apoptosis induction. In conclusion, interaction of TNFR1 with TNFR2 determines the biological characters of HPSCC, and TNFR1 may dominate this process. Moreover, interaction between the two receptors plays important roles in determining the fates of HPSCC cells and thus may serve as a therapeutic target for developing new therapeutic strategies for HPSCC.

[Metformin inhibits the proliferation of hypopharyngeal carcinoma Fadu cells and enhances the chemotherapeutic sensitivity of cells].

OBJECTIVE: To investigate the role of metformin on the growth inhibition induced by chemotherapeutic agents in hypopharyngeal carcinoma Fadu cells. METHODS: Fadu cells were treated with different concentrations of metformin for different time or treated with different concentrations of cisplatin, 5-fluorouracil or paclitaxel with or without metformin 5 mmol/L. MTT assay was used to evaluate the influence of metformin on the proliferation of Fadu cells. Cell-cycle was analyzed by flow cytometry. The expressions of AMP-dependent/activated protein kinase (AMPK) and P21 were examined by immunocytochemistry. RESULTS: Metformin inhibited the proliferation of Fadu cells in a dose-and time-dependent manner.Flow cytometry showed that cell cycle arrest in G1 phase was induced by metformin in Fadu cells.Immunocytochemistry showed the expressions of both AMPK and P21 in cells treated with metformin were higher than those in cells untreated with metformin. The growth inhibition of cells induced by cisplatin or paclitaxel but not 5-fluorouracil was enhanced by metformin. The combined indexes of cisplatin/paclitaxel/5-fluorouracil and metformin for 48 h were 0.43, 0.37, and 1.15, respectively. CONCLUSIONS: Metformin may inhibit the proliferation of Fadu cells by inducing the cell cycle arrest in G1 phase mediated in part by AMPK and P21. Metformin enhances the sensitivity of Fadu cells to cisplatin and paclitaxel.

Knockdown of glucose-regulated protein 78 abrogates chemoresistance of hypopharyngeal carcinoma cells to cisplatin induced by unfolded protein in response to severe hypoxia.

Hypoxia renders tumor cells with reduced sensitivity and increased resistance to chemotherapeutic agents. One of the possible mechanisms underlying this unfavorable status is activation of the unfolded protein response (UPR) under hypoxic conditions, due to the upregulation of glucose-regulated protein 78 (GRP78) expression. GRP78, an endoplasmic reticulum chaperone protein and a key regulator of the UPR, has been reported to be overexpressed in various types of cancer. However, the role of GRP78 in regulating the cell growth and apoptosis of hypopharyngeal carcinoma cells, with regard to the severity of hypoxia, remains unclear. Therefore, the aim of the present study was to investigate whether, and under what circumstances, GRP78 is associated with hypoxia-induced chemoresistance in hypopharyngeal carcinoma. For this purpose, cells from the FaDu human hypopharyngeal carcinoma cell line were cultured under normoxic and hypoxic conditions for different time periods. No significant changes in GRP78 and C/EBP homology protein (CHOP) protein expression levels were revealed under moderately hypoxic conditions (oxygen concentration, 1%), but these levels were changed over time under severely hypoxic conditions (oxygen concentration, <0.02%). This indicated that severe hypoxia, rather than moderate hypoxia, leads to UPR activation in hypopharyngeal carcinoma cells. Knockdown of GRP78 with short hairpin RNA inhibited cell proliferation and promoted apoptosis under severely hypoxic conditions, even with cisplatin treatment, indicating that GRP78 confers FaDu cells resistant to chemotherapy in response to severe hypoxia. Furthermore, knockdown of GRP78 resulted in a significant increase in CHOP and Bax expression levels and a decrease in Bcl-2 expression levels with simultaneous increase in the levels of apoptosis under severely hypoxic conditions. It was concluded that severe hypoxia leads to UPR activation and elevation of GRP78 expression, promoting cell survival and inducing chemoresistance. Silencing of GRP78 may block the pro-survival arm of UPR, simultaneously promoting proapoptotic signaling through induction of CHOP. Downregulation of GRP78 may be a promising strategy for overcoming the resistance of hypopharyngeal cancer to chemotherapy.

Regulation of glucose transporter protein-1 and vascular endothelial growth factor by hypoxia inducible factor 1α under hypoxic conditions in Hep-2 human cells.

The present study evaluated the regulation of glucose transporter protein-1 (Glut-1) and vascular endothelial growth factor (VEGF) by hypoxia inducible factor 1α (HIF-1α) under hypoxic conditions in Hep-2 human cells to explore the feasibility of these three genes as tumor markers. Hep-2 cells were cultured under hypoxic and normoxic conditions for 6, 12, 24, 36 and 48 h. The proliferation of Hep-2 cells was evaluated using an MTT assay. The protein and mRNA expression levels of HIF-1α, Glut-1 and VEGF were detected using the S-P immunocytochemical method, western blotting and reverse transcription polymerase chain reaction (RT-PCR). The results revealed that the expression levels of HIF-1α, Glut-1 and VEGF protein in Hep-2 cells were significantly elevated under hypoxic conditions compared with those under normoxic conditions over 36 h. Under hypoxic conditions, mRNA levels of HIF-1α were stable, while mRNA levels of Glut-1 and VEGF changed over time. In conclusion, Glut-1 and VEGF were upregulated by HIF-1α under hypoxic conditions in a time-dependent manner in Hep-2 cells and their co-expression serves as a tumor marker.

[Impacts of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism].

OBJECTIVE: To investigate the effects of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism. METHODS: The shRNA interference recombinant plasmid targeting HIF-1α was synthesized and transfected into Hep-2 cells. The HIF-1α knockdown Hep-2 cells were established after clonal selection and the expression of HIF-1α was measured. The cellular features including proliferation, clonal formation, cell cycle, apoptosis and CD133 phenotype were measured in Hep-2 cells cultured under hypoxic condition in vitro. CD133+ cells were sorted from Hep-2 cells with flow cytometry. Clonal formation test and cisplatin treatment were carried out, and the expressions of related genes (Oct-4, suvivin and p53) in CD133+ cells were measured. RESULTS: HIF-1α knockdown Hep-2 cells was successfully established, as evidenced by the reduced mRNA and protein expressions of HIF-1α. The Hep-2 cells cultured under hypoxic microenvironment showed higher proliferation and clonal formation activity, cell cycle arrest in G0/G1, lower apoptosis, up-regulated CD133, however the effects of hypoxia reduced in HIF-1α knockdown Hep-2 cells. CD133+ cells were successfully sorted from Hep-2 cells, and the CD133+ cells showed increased clonal formation activity and cisplatin treatment resistance in hypoxia. Also the effects of hypoxia on CD133+ cells decreased with HIF-1α knockdown, showing down-regulated Oct-4 and survivin and up-regulated p53. CONCLUSIONS: Hypoixa can induce the features of cancer stem cells in Hep-2 cells and increase proliferation, differentiation and chemoresistant ability of CD133+ cells, which might be correlated with the changes in expressions of HIF-1α and related genes regulated by HIF-1α.

[Expression of hypoxia inducible factor-1α and correlated target genes in human laryngeal carcinoma].

OBJECTIVE: To detect the expression of hypoxia inducible factor 1 alpha (HIF-1α), glucose transporter protein-1 (GLUT-1) and vascular endothelial growth factor (VEGF) in human laryngeal carcinoma tissue, and to study the relationship between hypoxia and HIF-1α, GLUT-1, VEGF in human laryngeal carcinoma Hep-2 cells and to explore the effect of HIF-1α, GLUT-1 and VEGF as endogenous hypoxic markers on laryngeal carcinoma. METHODS: The expression levels of HIF-1α, GLUT-1 and VEGF were detected in 35 cases of laryngeal carcinoma by SP immunohistochemical methods and in Hep-2 cells by SP immunocytochemical methods. The relationship between HIF-1α and GLUT-1, VEGF protein expression was analyzed. RESULTS: Of the 35 cases, 16 cases expressed HIF-1α, 16 cases expressed GLUT-1, 19 cases expressed VEGF. The expression of HIF-1α and VEGF were closely correlated with pathologic grading and lymphnode metastasis. GLUT-1 was correlated with lymphnode metastasis. The expression levels of HIF-1α, GLUT-1 and VEGF in Hep-2 cells under hypoxic condition were higher than those under normoxic condition. CONCLUSION: HIF-1α may promote the expression of GLUT-1 and VEGF in laryngeal carcinoma, furthermore promote tumor angiogenesis, invasion, and metastasis of the laryngeal carcinoma.

Biological and clinical significance of p75NTR expression in laryngeal squamous epithelia and laryngocarcinoma.

CONCLUSION: The apparent features of p75 neurotrophin receptor (p75(NTR)) expression indicated that p75(NTR) would serve as a potential stem cell marker for normal human laryngeal squamous epithelia. In human laryngeal squamous cell carcinoma (LSCC) p75(NTR) is differentially expressed. The abnormal expression and distribution of p75(NTR) may indicate malignant transformation. OBJECTIVE: To investigate the expression of p75(NTR) and its possible roles in normal laryngeal squamous epithelia and LSCC. METHODS: We used immunohistochemistry methods to examine normal laryngeal epithelia, para-cancer mucosa with dysplasia, laryngeal papilloma, and LSCC specimens for the expression of p75(NTR), nerve growth factor (NGF), -tyrosine kinase receptor (TrkA), p63, and Ki67. Immunocytochemistry and flow cytometry were used to examine the expression of p75(NTR) in Hep-2 cells. RESULTS: The expression of p75(NTR) was only located in basal cells of normal laryngeal epithelia, consistent with the staining features of epithelial stem cells as evidenced by parallel staining of p63, a putative keratinocyte stem cell marker. p75(NTR) is differentially expressed in LSCC, although no significant relationship was found with many clinicopathologic factors, this expression and distribution may correlate to malignant transformation and tumor proliferation. Co-expression of p75(NTR) and CD133 was confirmed, showing the association of p75(NTR)-positive cells with cancer stem cells in Hep-2 cells.