Cellular States and Secondary Chemical Bonding: A Biochemical View of Major Human Diseases.

Nasopharyngeal carcinoma (NPC) is different from other head and neck squamous cell carcinomas and is closely related with Epstein-Barr virus infection. It is endemic in southern China and Southeast Asia, affecting between 20 and 30 per 100,000 populations. According to the World Health Organization (WHO) histological classification, there are three subtypes of NPC: WHO type 1 NPC is keratinizing squamous cell carcinoma; type 2 NPC is differentiated non-keratinizing carcinoma; type 3 NPC is undifferentiated non-keratinizing carcinoma. In southern China including Hong Kong, type 3 NPC (undifferentiated NPC) is dominant and constitutes over 90% of the total NPC. MicroRNA-138 (miR-138) is a small non-coding RNA which has been reported to be highly expressed in undifferentiated NPC. We hereby evaluated whether the miR-138 level could be used to differentiate NPC patients from the normal individuals and examine the potential oncogenic role in undifferentiated NPC cell line. To validate the hypothesis that miR-138 was an oncogenic microRNA, which is overexpressed in undifferentiated NPC patients, we first examined its expression level in nasopharyngeal tissues and peripheral blood. In our cohort, cancer tissues samples were collected from 42 primary NPC and 29 recurrent NPC patients. To evaluate the expression level in the cancer tissues, the miR-138 level was quantified by real-time quantitative polymerase chain reaction. For primary NPC, the expression level was compared with 29 normal nasopharyngeal epithelia. For recurrent NPC, the microRNA level was compared with the paired normal mucosa counterparts obtained from the same patients. In addition, plasma samples were also collected from 22 primary NPC, 21 recurrent NPC and 17 normal individuals. Our data suggested that there was no difference in the miR-138 expression level in primary NPC tissue and normal nasopharyngeal tissue from control. There was no difference in the circulating miR-138 levels in the primary NPC, recurrent NPC and normal control groups. The circulating miR-138 could not be used to differentiate NPC patients from the normal individuals. Further functional analysis on the undifferentiated NPC cell line HONE1 suggested that miR-138 overexpression could enhance NPC cell proliferation, migration and invasion in comparison with the mock control. With the use of high-throughput gene expression arrays, we observed that multiple cancer-related pathways were affected in miR-138 overexpressed NPC cells. Staining with Acridine orange (AO) and phosphorylated H2AX (γH2AX) showed that miR-138 overexpression is associated with an enhanced response to radiation. Our results are concordant with other similar studies suggested that miR-138 is an oncogenic microRNA which play an important part in the undifferentiated NPC tumorigenesis. Further studies, based on larger sample size, are warranted to explore the clinical use of this small RNA in diagnosis, prognosis and management of undifferentiated NPC.

NOLC1, an Enhancer of Nasopharyngeal Carcinoma Progression, Is Essential for TP53 to Regulate MDM2 Expression

Temporal-spatial patterns of Mesozoic Paleo-Pacific and Tethyan supra-subduction systems in SE Asia: Key observations and controversies in Borneo and its surroundings

Nasopharyngeal carcinoma is one of the most common types of malignant tumor in Southern China and Southeast Asia, and its etiology is closely associated with Epstein-Barr virus (EBV) infection. Non-keratinizing carcinoma accounts for >95% of all nasopharyngeal carcinoma cases. In addition, metastatic nasopharyngeal carcinoma from other locations in the body is extremely rare. This study reports the case of a 53-year-old female who presented with a lesion on the left nasal alar skin that had slowly developed over a five-year period. A biopsy was obtained and the lesion was histologically diagnosed as cutaneous squamous cell carcinoma (SCC). A nasopharyngeal neoplasm was also detected by 18-fluorine-2-fluoro-2-deoxy-d-glucose positron emission tomography/computed tomography and nasopharyngoscopy. A biopsy of the nasopharyngeal neoplasm confirmed a diagnosis of SCC. However, a small EBV-encoded nuclear RNA (EBER) test demonstrated that the nasopharyngeal tumor cells were all negative for EBV. As the majority of nasopharyngeal carcinomas were positive for EBER, it was concluded that the nasopharyngeal carcinoma had metastasized from the cutaneous SCC. A brief review of literature is also presented, in addition to a discussion of the pathogen, epidemiology and diagnosis of cutaneous and nasopharyngeal carcinomas.

Nasopharyngeal carcinoma (NPC) is prevalent in southeastern Asia, and its tumorigenesis is rather complex. The purpose of this research was to identify the pivotal genes that may be altered during the early stage of NPC progression. Eleven genes were selected by comparative microarray analysis of NPC versus normal nasomucosal cells. The expression of SPARC (secreted protein, acidic, cysteine-rich) was statistically significantly downregulated in NPC cells. In exploring the mechanism underlying the decreased transcription of SPARC in NPC cells, we found that the transcription factor SRY (sex-determining region Y)-box 5 (SOX-5) is upregulated in NPC cells. RNA interference of SOX-5 by short hairpin RNA (shRNA) in NPC cells caused a dramatic increase in SPARC and chromosome immunoprecipitation assay showed SOX-5 can bind directly to SPARC promoter, suggesting that SOX-5 acts as a key transcriptional repressor of SPARC. We further demonstrate that shRNA knockdown of SOX-5 suppressed the proliferation of NPC cells, as well as their migratory ability, which was also observed when SPARC was overexpressed in NPC cells. Alternatively, blocking SPARC with an antagonistic antibody reversed the effects of SOX-5 knockdown. In 66 NPC patients’ biopsy specimens, overexpression of SOX-5 in tumor cells correlated clinically with poor survival. Our study suggests that SOX-5 transcriptionally downregulates SPARC expression and plays an important role in regulation of NPC progression. SOX-5 is a potential tumor marker for advanced NPC prognosis.

Nasopharyngeal carcinoma (NPC) is an endemic tumor in southern China and Southeast Asia, as well as in Taiwan. The main etiologic factors include genetic susceptibility, chemical carcinogens and association with Epstein-Barr virus (EBV) infection. NPC is highly radiosensitive and chemosensitive. Presently, radiotherapy remains a common treatment for early disease, while concurrent chemoradiotherapy is being increasingly accepted as the standard treatment for advanced disease. Although NPC at early stages can be cured by radical radiotherapy, there is a high recurrence rate in patients with advanced NPC. NPC is a highly metastatic and invasive malignant tumor in which the EBV genes encoding LMP-1 is expressed. Latent membrane protein 1 (LMP1), an EBV membrane protein is considered to be the EBV oncoprotein. Matrix metalloproteinase (MMP) -2, -9 are the MMP families, degrade Type IV collagen, a major component of extracellular matrix and is believed to be crucial for cancer invasion and metastasis. Recently, the study has shown that LMP1 induces MMP-9 in vitro cell line, which suggests the possibility of a mechanism in which LMP1 of EBV contributes to the invasion and metastasis of NPC by the induction of MMP-9. Upregulation of MMP-2 and -9 expression is observed in many cancers and high level of these proteins are found in peripheral blood of many cancer patients and increased plasma level of MMP-9 has been found in lung and breast and rectal cancer post radiotherapy. In this study, we aimed at evaluating the plasma pro-MMP-2 and pro-MMP-9 pro-enzymes levels during the course of CCRT and their clinical significances in patients with nasopharyngeal carcinoma. Patients with histological confirmed NPC attending the Department of Radiation Oncology at the Chung Chan Medical University Hospital. Cancer stage is according to 2002 AJCC staging system. All patients were treated with a concurrent chemoradiotherapy protocol. All patients were drawn 5mL of peripheral blood into an EDTA blood collection tubes with four times totally. The plasma pro-MMP2 and pro-MMP9 activity were measured in 20 NPC patients by gelatin zymography and the level were measured by enzyme linked immunosorbant assay (ELISA). The study result showed the plasma MMP-2,-9 concentration were not correlated with sex, age, T stage, lymph node metastatsis. But the neutrophils count was positive correlation to the plasma MMP-9 concentration that were statistically significant. In addition, the plasma MMP-9 levels ( mean= 68.6 ng/ml ) in the advanced stage are higher than in early stage ( mean= 45.7 ng/ml ) patients, while no difference in plasma MMP-2 levels was found. Furthermore, the authors have also find the kinetic of plasma MMP-9 concentration during CCRT in Group 2. was different from Group 1. NPC patients. The plasma MMP-9 concentration was still at high level in Goup 2. patients post CCRT. Our results were not statistically significant, because the sample sizes were small. Our conclusion, CCRT can be reduced effectively the plasma MMP-9 levels in early stage NPC. The more aggressive NPC patients still have high level of plasma MMP-9 post combine neo-adjuvant with CCRT. A high level of plasma MMP-9 may facilitate spreading of cancer cell and therefore a more aggressive tumor dissemination, recurrence and distant metastatsis.

Nasopharyngeal carcinoma (NPC) is prevalent in southeastern Asia, particularly southern China, Singapore and Taiwan. The aim of this study was to identify the pivotal genes that may be altered during NPC progression. Using cDNA microarray analysis, we compared the expression of 18 genes between NPC and normal nasomucosal cells. Q-RT-PCR analysis found the expression of IBFBP-6 in NPC cell lines and immunolocalization of IGFBP-6 in NPC to be very weak. To explore the effects of IGFBP-6 on NPC tumour growth, we constructed inducible plasmids containing full length of IGFBP-6 c-DNA (pBIG2i-IGFBP-6) and established pBIG2i-IGFBP-6 transfected NPC stable cell lines (NPC-TW01-pBIG2i-IGFBP-6). We then performed functional analysis of the IGFBP-6 in cell lines in vitro and in vivo. Overexpression of IGFBP-6 significantly suppressed the proliferation, invasion, and metastatic activity of NPC cells and increased their apoptosis. We found EGR-1, caspase-1 and TSP-1 genes to be markedly upregulated when NPC-pBIG2i-IGFBP-6 was treated with doxycycline. Knocking down EGR-1 with EGR-1 siRNA resulted in a decrease in expression of caspase-1, TSP-1 and EGR-1, but not the expression of IGFBP-6. However, in knockdown cells the unchanged expression of IGFBP-6 did not inhibit the migration of NPC cells. Chromatin immunoprecipitation and luciferase reporter assay experiments showed that IGFBP-6 bound the EGR-1 promoter regions and activated EGR-1 promoter. We conclude that IGFBP-6 can regulate the progression of NPC by regulating the expression of EGR-1. These results suggest that IGFBP-6 could be used as a new target in NPC therapy.

The transformation from hunter-gathering to farming in the south China coast has always been a conspicuous topic, as its great significance for the understanding of crop dispersal and human migration into southern China and Southeast Asia. It has been primarily assumed that rice was the only crop cultivated by early farmers in this region since 5,000 cal. BP., but the reliability of this speculation remains ambiguous, owing to the lack of systematic evidence. Based on analysis of macroscopic plant remains and phytoliths, as well as AMS radiocarbon dating at the Gancaoling site in Guangdong province, this study demonstrates the emergence of agriculture in the south China coast could be dated back to as early as 4,800–4,600 cal. BP., with the cultivation of rice and foxtail millet. This subsistence strategy change was an integral part of a more comprehensive social transformation, which started a new era of local history. Moreover, this discovery also provides further evidence supporting the universality of mixed farming in southern China and shed new light on the study of agriculture southward dispersal. The crop package of rice and millets possibly spread into the south China coast from Jiangxi via the mountain areas and then into Mainland Southeast Asia by a maritime route along the coastal regions.

Nasopharyngeal carcinoma (NPC) is a unique epithelial malignancy that occurs at a high frequency in certain regions of Southeast Asia. Previous study revealed the association between Epstein Barr Virus (EBV) and to a lesser extent, Human Papilloma Virus (HPV) with NPC. The role of the EBV in the pathogenesis of NPC was further supported by the recovery response of the tumor cells after being treated with EBV-specific T lymphocytes. The present study aims to determine the frequency distribution of EBV infection among the pathologically confirmed NPC patients and healthy control in Surabaya, Indonesia. The presence of Human Papilloma. Virus infection (HPV) was also analysed. The results indicated that EBV DNA existed in the 88% of the nasopharyngeal tissue biopsies of 25 NPC patients and none in the 10 healthy controls, and the difference was statistically significant.Analysis of the mononuclear cells of peripheral blood revealed that 60% of the 25 NPC patients carried EBV DNA whereas in control group 20% was found to be positive and the difference was statistically significant. Although a high positive rate EBV-DNA was detected in NPC patients, additional environmental and genetic factors must still be considered. Nevertheless, no HPV-DNA was detected from mononuclear cells of peripheral blood and nasopharyngeal tissue of the two groups. By this study there was no coexistence between the infection of EBV and HPV which promote carcinogenesis in NPC. Analysis using LMP1-DNA in tumor cell biopsies indicated that 72% of the NPC patients yielded PCR products and none of the healthy control, the difference was statistically significant. In conclusion the findings confirm the tight association between the EBV and NPC in Indonesia and that the specific presence of LMP1-DNAs in the tumor cells strongly indicates the important role of EBV in the pathogenesis of NPC. Coexistence of EBV and HPV infections was not found in NPC cases in Indonesia.

Environmental exposures, particularly infection with Epstein-Barr virus (EBV) and tobacco, are known risk factors for oral cancer. Studies in migrants may provide valuable insight into the environmental and genetic etiology of cancer. We wanted to define nasopharyngeal and hypopharyngeal carcinoma among immigrants in Sweden. The nationwide Swedish Family-Cancer Database (FCD) was used to calculate standardized incidence ratios (SIRs) for nasopharyngeal and hypopharyngeal carcinomas among the first-generation immigrants compared to the native Swedes. The FCD included 1969 and 691 cases of nasopharyngeal and hypopharyngeal carcinoma in the male and female Swedes and 178 and 65 cases in immigrants, respectively. The median age at diagnosis (years) was 63 among Swedes and 55 among immigrants. The risk of nasopharyngeal carcinoma was significantly higher in male (SIR = 35.6) and female (24.6) Southeast Asians, male (12.4) and female (34.7) North Africans, male (4.9) and female (10.9) Asian Arabs and some other male Asians immigrants (6.2 to 6.7). Among immigrants from European countries, only the men from former Yugoslavian showed an elevated risk (2.7). Hypopharyngeal carcinoma risk was only increased among the male immigrants from the Indian Subcontinent (5.4). Early life infection with EBV in countries of origin and probably a minor contribution by tobacco smoking may be the main environmental exposures influencing nasopharyngeal carcinoma risks among immigrants to Sweden. The high rates of hypopharyngeal carcinoma among Indian immigrants may point to a continued using of smokeless tobacco.

Nasopharyngeal carcinoma (NPC) is a common malignant cancer in Guangxi, as well as in southern China and Southeast Asia. NPC preferentially metastasizes to bone in advanced patients, major factor resulting in high mortality. The molecular basis of NPC development and cancer-induced bone lesions are still unclear. We and others have reported that chemokine, like CCL2, CXCL16, CXCL12, and their corresponding receptors played important roles in cancer bone metastasis. In this study, we firstly investigated that CCR2 and CXCR6 expressions in clinical specimens. Then, we measured CCL2, CCR2, CXCL16 and CXCR6 expression levels by RT-PCR, ELISA and western blot in NPC cell lines. Further, we used MEK inhibitor, U0126, to treat the NPC cell lines and found CCL2 and CXCL16 expressions significantly decreased. The NPC cell lines proliferation were inhibited by U0126 in dose- and time-dependently manner. Finally, we collected conditioned medium (CM) from the NPC cell lines absent or present U0126 treatments, to treat mouse bone marrow non-adherent cells in order to detect NPC CM-induced multinucleated osteoclast-like formation. The CMs collected from U0126 treated NPC cells were dramatically diminished the numbers of multinucleated osteoclasts. These results indicate that U0126 inhibits NPC cell growth and diminishes cancer-induced osteoclast formation partially through modulating the expression of CCL2 and CXCL16. Thus this study may provide a novel therapeutic strategy for advanced NPC patients, especially for treatment of NPC bone metastasis. Citation Format: Yu Zhu, Qiong Song, Yi Lu. MEK inhibitor diminishes nasopharyngeal carcinoma (NPC) cell growth and NPC-induced osteoclastogenesis via modulating CCL2 and CXCL16 expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4131. doi:10.1158/1538-7445.AM2015-4131

Nasopharyngeal carcinoma (NPC) is a malignant epithelial cell-derived tumor that is predominately found among populations in Southeast Asia, including Taiwan. The interaction that exists between tumors and the immune system is important in tumor progression. Because the mechanisms underlying NPC’s ability to escape the surveillance of the immune system are still unclear, our study sought to investigate the interaction between one type of antigen presenting cells, known as Dendritic cells (DCs), and NPC. DCs express a C-type lectin, DC-pecific ICAM-3-Grabbing Nonintegrin (DC-SIGN) that can induce suppressive immune responses by recognizing carbohydrate structure. Recent studies indicated that cancer cells express many abnormal glycosylated proteins, and these glycoproteins are involved in the immune suppressive responses. Therefore we used flow cytometry and immunostaining, and found that DC-SIGN-recognized ligands were expressed on NPC cells. Using enzyme-linked immunosorbent assay (ELISA), we demonstrated that NPC cells induced immunosuppressive interleukin (IL)-10 secretion from DC. We further showed that IL-10 secretion was inhibited by DC-SIGN antibody and siRNA. We immunoprecipitated the NPC membrane proteins using purified recombinant DC-SIGN protein, and identified the unknown proteins by LC/MS/MS. Annexin A2 was identified as a major protein which is highly expressed in TW01 NPC cells. In conclusion, NPC cells express DC-SIGN ligands, and stimulate immunosuppressive IL-10 secretion from DC through DC-SIGN activation, resulting in the escape from surveillance of the immune system. We are focusing on annexin A2, a novel target on nasopharyngeal carcinoma, and will research the relationship of annexin a2 and DC-SIGN in the future.

Among all head and neck (H&N) cancers, nasopharyngeal carcinoma (NPC) represents a distinct entity regarding epidemiology, clinical presentation, biological markers, carcinogenic risk factors, and prognostic factors. NPC is endemic in certain regions of the world, especially in Southeast Asia, and has a poor prognosis. In Indonesia, the recorded mean prevalence is 6.2/100 000, with 13 000 yearly new NPC cases, but otherwise little is documented on NPC in Indonesia. Here, we report on a group of 1121 NPC patients diagnosed and treated at Dr. Cipto Mangunkusumo Hospital, Jakarta, Indonesia between 1996 and 2005. We studied NPC incidence among all H&N cancer cases (n=6000) observed in that period, focusing on age and gender distribution, the ethnic background of patients, and the disease etiology. We also analyzed most prevalent signs and symptoms and staging of NPC patients at first presentation. In this study population, NPC was the most frequent H&N cancer (28.4%), with a male-to-female ratio of 2.4, and was endemic in the Javanese population. Interestingly, NPC appeared to affect patients at a relatively young age (20% juvenile cases) without a bimodal age distribution. Mostly, NPC initiated in the fossa of Rosenmuller and spreaded intracranially or locally as a mass in the head. Occasionally, NPC developed at the submucosal level spreading outside the anatomic limits of the nasopharynx. At presentation, NPC associated with hearing problems, serous otitis media, tinnitus, nasal obstruction, anosmia, bleeding, difficulty in swallowing and dysphonia, and even eye symptoms with diplopia and pain. The initial diagnosis is difficult to make because early signs and symptoms of NPC are not specific to the disease. Early-age Epstein-Barr virus (EBV) infection combined with frequent exposure to environmental carcinogenic co-factors is suggested to cause NPC development. Undifferentiated NPC is the most frequent histological type and is closely associated with EBV. Expression of the EBV-encoded latent membrane protein 1(LMP1) Oncogene in biopsy material was compared between NPC patients of < 30 years old and those of ≥ 30 years old, matched for sex and tumor stage. Higher LMP1 expression in patients of <30 years old was observed, which was related to more locoregional progressivity. Increased medical awareness of prevailing early stage signs and symptoms coupled to use of EBV-related diagnostic tumor markers may lead to down-staging and timely treatment to improve survival of patients with this aggressive disease.

Nasopharyngeal carcinoma (NPC) is a malignancy of the head and neck that is prevalent in Southeast Asia and southern China. Recent studies in epigenetics suggest that DNA methylation plays a pivotal role in the onset and progression of cancer. Combining the methyl-DNA binding domain capture technique and cDNA microarray analysis, we identified a unique hypermethylated gene, RERG (Ras-like estrogen-regulated growth inhibitor), that was down-regulated in NPC tissues. RERG is a tumor suppressor gene that was first reported in breast cancer. However, the functions of RERG are largely unknown in other tumor types. RERG expression was assessed in human subjects (NPC primary tissues and non-cancer tissues) and cell lines (NPC cell lines and an immortalized epithelial cell line NP460). Further, we investigated the methylation rate of RERG in both human subject and cell lines. 5-Aza-2’-deoxycytidine (Aza) or combined with trichostatin A (TSA) were treated to three NPC cell lines (HK1, C666-1 and HK1_EBV). In addition, the role of RERG in NPC cells and its underlying mechanisms were explored by overexpression of RERG in NPC cell lines. RERG was significantly down-regulated in NPC cancer nests compared to normal nasopharyngeal epithelium cells. Furthermore, the RERG promoter was frequently methylated in NPC tissues and cell lines. The RERG methylation rate yielded an area under the curve (AUC) of receiver operating characteristic (ROC) curve was 0.897 (95%CI: 0.818–0.976). The down-regulation of RERG was restored in NPC cells treated with Aza and TSA. In addition, ectopic expression of RERG in NPC cell lines resulted in a significant suppression of cell proliferation, clonogenicity, migration and invasion. RERG-overexpressing cells showed significantly slower growth and less angiogenesis in tumor xenografts in nude mice. RERG suppressed the ERK/NF-κB signaling pathway and inhibited tumor growth and angiogenesis with down-regulation of MMPs and IL8 in tumors of nude mouse xenografts. Our results suggest that RERG is frequently silenced by promoter CpG methylation in NPC, and acts as a functional tumor suppressor by suppressing the ERK/NF-κB signaling pathway. These findings support the potential use of RERG as a novel molecular target in NPC therapy.

BackgroundNPC is a highly invasive squamous-differentiated carcinoma, more common in east and southeast Asia, and the Epstein-Barr virus (EBV) plays a key role in its pathogenesis. Recurrent and metastatic NPC...