Erratum: Zinc-α2-glycoprotein 1 promotes EMT in colorectal cancer by filamin A mediated focal adhesion pathway: Erratum.

[This corrects the article DOI: 10.7150/jca.35380.].

LncRNA EPB41L4A-AS1 Regulates Cell Proliferation, Apoptosis and Metastasis in Breast Cancer.

OBJECTIVE: Breast cancer is a leading global public health problem. In our previous study, we identified that lncRNA EPB41L4A antisense RNA 1 (EPB41L4A-AS1) was significantly downregulated in breast cancer. However, the functional role of EPB41L4A-AS1 in breast cancer has not been clarified. Here, we further confirmed the expression and biological function of EPB41L4A-AS1 in breast cancer. MATERIALS: To demonstrate the role of EPB41L4A-AS1 in breast cancer, we transfected breast cancer lines with pcDNA3.1-EPB41L4A-AS1 expression vector to induce ectopic overexpression of EPB41L4A-AS1. Then, to explore the role of EPB41L4A-AS1 overexpression in breast cancer cell growth, cell cycle, apoptosis, invasion, and migration capacity, we performed CCK-8 assay, colony formation assay, flow cytometry analysis, wound recovery and transwell assay, respectively. We also constructed a co-expression network to explore the potential effect mechanism of EPB41L4A-AS1. RESULTS: Our research showed EPB41L4A-AS1 expression was significantly lower in tumor tissues than in adjacent non-cancerous tissues. Overexpression of EPB41L4A-AS1 significantly reduced the proliferation of breast cancer cells. Flow cytometric analysis showed that forced expression of EPB41L4A-AS1 significantly increased the apoptosis rate of breast cancer cells. In addition, we found that upregulated EPB41L4A-AS1 significantly inhibited the migration and invasive ability of breast cancer. Functional analysis of co-expressed mRNAs suggested that EPB41L4A-AS1 may be involved in ribosomal, cell cycle, spliceosomal and p53 signaling pathways. CONCLUSION: Our findings suggest that EPB41L4A-AS1 is a tumor suppressor gene in breast cancer.

Nasopharyngeal metastasis from colorectal cancer: a case report.

Metastases from colorectal cancer can occur either through the lymphatic or by hematogenous spread. The most common metastatic sites are the lung and liver. Nasopharyngeal metastasis from colorectal cancer has never been previously reported in the literature on the internet databases we can found. In this paper, we present the case of a 79-year-old male suffering from adenocarcinoma of the rectum with distant metastases to the liver, lung, and nasopharynx. Over the previous 7 years, he had received treatment for rectal cancer including radical surgery (miles surgery), chemotherapy, hepatectomy, and pneumonectomy. After local nasopharyngeal radiotherapy, the patient's quality of life significantly declined and they eventually died of dyspnea caused by airway obstruction due to a nasopharyngeal mass after 7 months of palliative treatment involving pain relief from end-stage disease. Nasopharyngeal metastases from colorectal cancer are extremely rare in the clinic. To the best of our knowledge, this is the first case reporting this occurrence which not only extends the disease database but also warns doctors to pay more attention to these clinical scenarios. Strict monitoring of patients with colorectal cancer after primary treatment could lead to the early diagnosis of metastases and give patients better opportunities for treatment and improved prognosis.

Zinc-α2-glycoprotein 1 promotes EMT in colorectal cancer by filamin A mediated focal adhesion pathway.

Liver metastasis is the main reason for the poor prognosis of colorectal cancer, and identifying molecules involved in liver metastases of colorectal cancer may provide effective therapeutic targets. Zinc-α2-glycoprotein 1(AZGP1) is a candidate biomarker for diagnosis and prognosis in cancer. However, its function and molecular mechanism in metastatic colorectal cancer remains largely unknown. We previously found that up-regulated AZGP1 promotes proliferation, migration and invasion in colorectal cancer cell line, here we elucidated the mechanism of AZGP1 in regulating metastasis. In this article, we found that AZGP1 was also highly expressed in colorectal cancer tissues with liver metastasis relative to those without metastasis, and abundant expression of AZGP1 was associated with poor prognosis, also, AZGP1 down regulation prevented cell metastasis in vivo and in vitro. We further demonstrated that AZGP1 promotes metastasis by regulating the epithelial-mesenchymal transition (EMT) and associating with molecules involved in the focal adhesion pathway, including the adhesion molecule FLNA, which acts as an important protein interactor. More importantly, AZGP1 down regulation inhibited the phosphorylation of FLNA mediated by the restrain of PAK2 kinase, thereby inducing its proteolysis and subsequently affecting its subcellular localization, where it regulates the EMT and promotes metastasis. Collectively, these results highlight AZGP1 as a new and promising therapeutic molecule for liver metastatic colorectal cancer.

The lncRNA UNC5B-AS1 promotes proliferation, migration, and invasion in papillary thyroid cancer cell lines.

The incidence of thyroid cancer detection is continually improving worldwide with the spread of diagnostic imaging and surveillance. Although we have made great progress, there are still unknown mechanisms of papillary thyroid cancer. We found that UNC5B-AS1 is a potential oncogene in thyroid cancer. Therefore, our study aimed to investigate the biological functions of the lncRNA UNC5B-AS1 in papillary thyroid cancer. As a result, RNA-seq data on primary papillary thyroid cancer (PTC) in the TCGA database were obtained. RT-qPCR was performed to evaluate the expression levels in thyroid tissue. We then analysed the expression level of UNC5B-AS1 and its association with clinicopathologic characteristics in the TCGA database. We downregulated UNC5B-AS1 using small interfering RNA and carried out assays of cell proliferation, colony formation, migration and invasion to explore the function of UNC5B-AS1 in PTC cell lines (TPC1 and BCPAP). These results suggested that the lncRNA UNC5B-AS1 was significantly upregulated in both the TCGA cohort and our tissue cohort. Upregulated UNC5B-AS1 correlated with lymph node metastasis (P < 0.001), tumor size (P = 0.002) and histological type (P = 0.013). We also achieved an area under the ROC curve (AUC) of 93.2% for our validated cohort, which was consistent with the AUC of 94.5% for the TCGA cohort, for differentiating between PTC tissues and normal tissues. Downregulating UNC5B-AS1 expression at the RNA level significantly inhibited cell proliferation, colony formation, migration, and invasion in PTC cell lines (TPC1 and BCPAP). This study demonstrated that the lncRNA UNC5B-AS1 plays an important role in tumourigenesis and metastasis of PTC and may be a potential therapeutic target for PTC.

KLP-PI: a new prognostic index for luminal B HER-2-negative breast cancer.

Luminal B HER-2-negative (LBHN) subtype is one of the major subtypes of breast cancer according to different features, clinical behaviors, and treatment response. The LBHN subtype shows a poor prognosis and is insensitive to endocrine therapy. Our work aim is to investigate the prognostic factor in the LBHN subgroup and, meanwhile, try to obtain an optimal prognostic index (PI) contrapose LBHN subgroup which helps to guide chemotherapy. A total of 515 female LBNH patients who underwent diagnosis and surgery at our hospitals from August 2008 to August 2018 were enrolled. Clinical-pathological information was obtained and immunohistochemistry result was available. From these cases, a 30% Ki-67 LI was employed to divide LBHN into two groups with low and high levels; high Ki-67 LI was associated with GIII tumor grade (P < 0.001), positive axillary lymph nodes (ALN) status (P = 0.018) and negative PR status (P = 0.016), and also seemed to be related to T2-T3 tumor size (P = 0.058). High Ki-67 level (HR = 3.30; P < 0.011), positive ALN (HR = 7.29; P < 0.001) and PR negative (HR = 2.63; P = 0.034) significantly associated with poor 5-year DFS in multivariate Cox's proportional hazard regression model. A novel prognosis prediction model (KLP-PI), based on Ki-67 LI, ALN and PR status, showed a better discriminatory ability compared with traditional Nottingham prognostic index targeted to LBHN breast cancer. Our study highlights that high Ki-67 LI, positive ALN and negative PR status were associated with poor outcome in LBHN patients, and composed by these prognostic factors, KLP-PI improves the prognostic assessment using the Nottingham Prognostic Index when aiming at LBHN subtype.

MAL2 promotes proliferation, migration, and invasion through regulating epithelial-mesenchymal transition in breast cancer cell lines.

BACKGROUND: Breast cancer is one of the most common malignant tumors in women. However, the underlying molecular mechanisms of breast cancer are still far to clear. With the development of sequencing technology, we discovered that MAL2 is overexpressed in tumor tissues. But the major function of MAL2 in breast cancer has not to be well confirmed. MATERIALS AND METHODS: We downloaded and analyzed the MAL2 expression in The Cancer Genome Atlas (TCGA) database. Real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to detect the expression of MAL2 in 35 breast cancer patients. Then, we performed proliferation, colony formation, migration, invasion and western blot assays to investigate the role of MAL2 in breast cancer cell lines (MDA-MB-231 and BT-549). RESULTS: In our research, we found that MAL2 is remarkably overexpressed in breast cancer tissues compared to adjacent non-cancer tissues by RT-qPCR (T: N = 5.28 ±â€¯4.34:1.82 ±â€¯1.11, P < 0.001) and high expression of MAL2 has worse overall survival in TCGA cohort (P = 0.0032). Knocked down MAL2 could decrease the ability of proliferation, migration, and invasion of breast cancer cell lines. Our Western Blot assay results investigated that MAL2 could regulate EMT. CONCLUSION: In this study, we demonstrated the function of MAL2 in breast cancer cell lines and it might act as an oncogene in breast cancer.

Upregulated LAMB3 increases proliferation and metastasis in thyroid cancer.

BACKGROUND: Thyroid cancer is the most commonly reported endocrine malignancy, and its increased incidence has been the highest in all human tumors in recent decades. To investigate the mechanism of papillary thyroid cancer (PTC) occurrence and progression, we performed RNA sequencing and found an upregulated gene, LAMB3. However, the biological function of LAMB3 is still not clear. MATERIALS AND METHODS: We analyzed LAMB3 expression using The Cancer Genome Atlas (TCGA) database and hypothesized LAMB3 to be a gene associated with PTC. To test this hypothesis, we collected 89 pairs of thyroid nodules and adjacent normal thyroid tissues (56 pairs of PTCs, 33 pairs of benign thyroid nodules). Afterward, we performed real-time quantitative polymerase chain reaction (RT-qPCR) to investigate LAMB3 expression in thyroid nodule patients, and then analyzed clinicopathologic features. We performed proliferation, colony formation, migration, and invasion assays to determine the function of LAMB3 in PTC. RESULTS: We demonstrated that LAMB3 plays oncogenic roles in PTC. The relative expression of LAMB3 is significantly upregulated in PTC compared with matched thyroid normal tissues in validated cohort and TCGA cohort (P<0.001). We also checked area under the curve (AUC of receiver operator characteristic [ROC]) of 97.3% for validated cohort and 90.1% for TCGA cohort to differentiate PTC tumors from normal tissues. In clinicopathologic feature analysis, we found that upregulated LAMB3 is closely related to lymph node metastasis (P=0.018). Furthermore, knockdown of LAMB3 inhibited the proliferation, colony formation, migration, and invasive capacity of PTC. CONCLUSION: This study indicated that LAMB3 is a gene associated with PTC.

Novel strategy of stents in thyroid mass: a case series report of managing severely dyspneic patients.

BACKGROUND: Tracheal and bronchial stenosis is a life-threatening condition causing difficulty in breathing and even severe respiratory distress. The silicone tracheobronchial stents were placed using the rigid bronchoscopy into the trachea of severe dyspneic patients and they exhibited symptomatic improvement as well as a rise in the saturation of oxygen. The bronchial stents were applicable to many extensive malignant airway stenosis patients, such as those with esophageal cancer, lung cancer, and laryngeal cancer. But the effectiveness of bronchial stents for thyroid cancer is not certain. CASE PRESENTATION: Here, we report 3 emergency patients with a thyroid mass referred to our hospital because of grade 4 dyspnea according to the American Thoracic Society shortness of breath guidelines. The main clinical symptoms were severe dyspnea and stridor. The radiographic examination and tomographic examination showed the narrowing and displacement of the trachea. To the best of our knowledge, ideal airway management for the massive thyroid mass was considered to be temporary tracheobronchial stent placement pre-operation. CONCLUSION: In our study, we applied the tracheobronchial stent to massive thyroid mass patients with dyspnea and aimed to not only improve preoperative airway obstruction but also to protect the potential airway collapse from post-operative tracheomalacia following extubation. We found that application of tracheobronchial stents may provide a new strategy to dyspneic patients with huge thyroid mass.

CDKL1 promotes tumor proliferation and invasion in colorectal cancer.

BACKGROUND: CDKL1 is a member of the cell division cycle 2 (CDC2)-related serine threonine protein kinase family and is overexpressed in malignant tumors such as melanoma, breast cancer, and gastric cancer. OBJECTIVE: This study aimed to evaluate whether CDKL1 can serve as a potential molecular target for colorectal cancer therapy. MATERIALS AND METHODS: Expression of CDKL1 in colorectal cancer tissues and cell lines was measured by immunohistochemistry and Western blot, respectively. To investigate the role of CDKL1 in colorectal cancer, CDKL1-small hairpin RNA-expressing lentivirus was constructed and infected into HCT116 and Caco2 cells. The effects of RNA interference (RNAi)-mediated CDKL1 downregulation on cell proliferation and invasion were assessed by CCK-8, colony formation, transwell, and tumorigenicity assays in nude mice. The effects of CDKL1 downregulation on cell cycle and apoptosis were analyzed by flow cytometry. Furthermore, microarray method and data analysis elucidated the molecular mechanisms underlying the phenomenon. RESULTS: CDKL1 protein was overexpressed in colorectal cancer tissues compared with paired normal tissues. Knockdown of CDKL1 in HCT116 and Caco2 significantly inhibited cell growth, colony formation ability, tumor invasion, and G1-S phase transition of the cell cycle. The knockdown of CDKL1 stimulated the upregulation of p15 and retinoblastoma protein. CONCLUSION: CDKL1 plays a vital role in tumor proliferation and invasion in colorectal cancer in vitro and in vivo and, thus, may be considered as a valuable target for therapeutic intervention.

Identification of lncRNA FAM83H-AS1 as a novel prognostic marker in luminal subtype breast cancer.

BACKGROUND: Luminal subtype breast cancer accounts for a predominant number of breast cancers. Considering the heterogeneity of the disease, it is urgent to develop novel biomarkers to improve risk stratification and optimize therapy choices. Long non-coding RNA (lncRNA) represents an emerging and understudied class of transcripts that play a significant role in cancer biology. Growing knowledge of cancer-associated lncRNAs contributes to the development of molecular markers for prognosis evaluation and gene therapy. MATERIALS AND METHODS: Three pairs of primary luminal subtype breast cancer tissues and adjacent non-cancerous tissues were collected and sequenced. EBseq algorithm was used to identify differentially expressed lncRNAs. RNA sequencing data from The Cancer Genome Atlas (TCGA) database were used to validate the robustness of our RNA-seq results. Kaplan-Meier and Cox regression analyses were utilized to assess the association between the lncRNAs and overall survival of patients in TCGA cohort. RESULTS: A total of 796 lncRNAs were significantly dysregulated in luminal subtype breast cancer, including 436 upregulated and 360 downregulated lncRNAs. Among them, FAM83H antisense RNA 1 (FAM83H-AS1) was the most upregulated lncRNA, whereas GSN antisense RNA 1 (GSN-AS1) was the most downregulated lncRNA. Moreover, we proved that the high expression level of FAM83H-AS1 indicated unfavorable prognosis not only in luminal subtype breast cancer but also in all subtype breast cancers. To the best of our knowledge, this is the first report indicating that FAM83H-AS1 was involved in luminal subtype breast cancer and was an independent prognostic indicator. CONCLUSION: Our study provides a rich resource to the research community for further identifying lncRNAs with diagnostic and therapeutic potentials and exploring biological function of lncRNAs in luminal subtype breast cancer.

Downregulation of paraoxonase 3 contributes to aggressive human hepatocellular carcinoma progression and associates with poor prognosis.

Paraoxonase (PON) enzymes possess antioxidant properties and protect against cardiovascular diseases. As a member of PON family, PON3 is primarily synthesized in the liver and poorly investigated. This study aimed to examine the expression of PON3 in human hepatocellular carcinoma (HCC) and investigate the clinical significance and biological function of PON3 in HCC patients. We first analyzed PON3 expression in 50 paired HCC samples (HCC tissues vs matched para-cancerous tissues) and 160 clinical HCC specimens by using immunohistochemistry (IHC). Our results showed that the expression of PON3 was downregulated in HCC and significantly associated with tumor-node-metastasis (TNM) stage, tumor size, and tumor number. Kaplan-Meier survival and Cox regression analyses showed that PON3 was an independent prognostic factor for overall survival (OS) and time to recurrence (TTR). Finally, we aimed to reveal the biological function of PON3 in HCC growth and metastasis, and our results showed that overexpression of PON3 potently inhibited growth and metastasis of HCC. Collectively, our study demonstrated that PON3 exhibited tumor-suppressive effects toward HCC and it might serve as a novel prognostic marker in HCC.

Downregulated long non-coding RNA CLMAT3 promotes the proliferation of colorectal cancer cells by targeting regulators of the cell cycle pathway.

Over-expression of long non-coding RNA (lncRNA)-CLMAT3 is significantly associated with colorectal liver metastasis and is an independent predictor of poor survival for colorectal cancer patients. However, as little is known regarding the role of this gene in the proliferation of colorectal cancer in vitro, we investigated the involvement of lncRNA-CLMAT3 in colorectal cancer cell proliferation. In this study, we demonstrate that lncRNA-CLMAT3 expression was significantly increased in colorectal cancer cells compared with a normal intestinal mucous cell line and that inhibition of lncRNA-CLMAT3 suppressed colorectal cancer cell proliferation in vitro. We also found that this reduced colorectal cancer cell proliferation due to lncRNA-CLMAT3 knockdown is associated with G0/G1 cell-cycle arrest induction and apoptosis enhancement. Furthermore, lncRNA-CLMAT3 knockdown enhanced Cdh1 expression and resulted in p27Kip accumulation via increased Skp2 protein ubiquitination. Taken together, our findings suggest that reducing lncRNA-CLMAT3 inhibits colorectal cancer cell proliferation by affecting cell cycle components.

Aberrant expression of long noncoding RNAs in colorectal cancer with liver metastasis.

Long noncoding RNA (lncRNA) plays a crucial role in the regulation of various cellular processes and human diseases. However, little is known about the role of lncRNAs in colorectal liver metastasis (CLM). In the present study, we aimed to determine whether lncRNAs are differentially expressed in CLM tissue and to further assess their clinical value. lncRNA arrays were employed to screen for differentially expressed lncRNAs in colorectal cancer (CRC) tissues with synchronous, metachronous, or nonliver metastasis. Based on bioinformatics data, a quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assay was performed to identify target lncRNAs in an expanded set of CRC samples with various subtypes of liver metastasis. The relationships between the target lncRNAs and the clinical characteristics and patient prognosis were further analyzed. After determining the expression profile of lncRNAs (n = 1332) in CLM tissue, 40 differentially expressed lncRNAs that were potentially related to CLM were selected for further examination in an expanded set of clinical samples, and three novel target lncRNAs, termed lncRNA-CLMAT1-3, were verified. High lncRNA-CLMAT3 expression strongly correlated with liver metastasis (P = 0.03) and lymph node metastasis (P = 0.009). Moreover, patients displaying high lncRNA-CLMAT3 expression exhibited a shorter median overall survival duration than those displaying low lncRNA-CLMAT3 expression (30.7 vs. 35.2 months, P = 0.007). Multivariate analysis demonstrated that the lncRNA-CLMAT3 expression level is an independent prognostic factor (hazard ratio 2.05, P = 0.02) after adjusting for other known prognostic factors. lncRNA-CLMAT3 over-expression was significantly associated with CLM and was an independent predictor of poor survival for patients with CRC.

[A retrospective survival analysis of with pulmonary metastasis from colorectal cancer].

OBJECTIVE: To evaluate the survival rate after pulmonary resection for metastatic colorectal cancer(CRC). METHODS: Clinical data of 77 patients with pulmonary metastasis from CRC between January 2005 and October 2008 in the Zhongshan Hospital, Fudan University were retrospectively analyzed. RESULTS: There were 38 patients with synchronous pulmonary metastasis, of whom 2 underwent resection for pulmonary metastasis. The median survival time of two groups was 25 months and 18 months, which was not significantly different (P=0.33). There were 39 cases of metachronous pulmonary metastasis, of whom 28 received pulmonary metastasis resection. The 1-year and 3-year survival rates of 2 groups were 93.3% and 58.5%, and 38.8% and 19.1%, respectively. The median survival time of two groups was 26.7 months and 8 months, and the difference was statistically significant (P=0.004). CONCLUSION: Surgical resection can improve the survival rate in patients with pulmonary metastasis from colorectal cancer.

[Prognostic analysis of 669 liver metastasis of colorectal cancer cases].

OBJECTIVE: To evaluate the relation between different therapy and survival rate of liver metastasis of colorectal cancer (LMCC). METHODS: Clinical data of 669 LMCC patients,collected from Fudan University Zhongshan Hospital from January 2000 to July 2008, were analyzed retrospectively. RESULTS: Of the 669 cases, 379 cases were synchronous liver metastases(SLM) and 290 cases were metachronous liver metastases(MLM). There were no significant differences in age, gender and position of primary tumor between SLM and MLM groups(P>0.05), but as to liver metastasis characteristics(liver lobe involved, focus number and maximal focus diameter) and CEA, CA19-9 before therapy,there were significant differences(P<0.05). Two hundred and fifty-three cases underwent curative hepatic resection, including 123 cases in SLM and 130 cases in MLM. Until October 31, 2008, all the cases were followed up. The median survival time of SLM was(11+/-1) months and of MLM(23+/-2) months(P<0.01). Five-year survival rate of SLM was 6.4% and of MLM 11.4%(P<0.01). As to different treatments, median survival time and 5-year survival rate of curative hepatic resection group were 37 months and 35.6%, and of non-operation groups(i.e. intervention, chemotherapy, radiofrequency therapy and percutaneous ethanol injection) were 5 to 26 months and 0 to 3.6% respectively(P<0.05). CONCLUSIONS: Curative hepatic resection is the first choice of liver metastasis of colorectal cancer, which can improve the survival rate. Resection rate and survival of MLM are better than those of SLM.