Dataset Information

KIF15 facilitates gastric cancer via enhancing proliferation, inhibiting apoptosis, and predict poor prognosis.

ABSTRACT:

Background

Kinesin superfamily proteins (KIFs) can transport membranous organelles and protein complexes in an ATP-dependent manner. Kinesin family member 15 (KIF15) is overexpressed in various cancers. However, the function of KIF15 in gastric cancer (GC) is still unclear.

Methods

GC patients' data from The Cancer Genome Atlas (TCGA) were analyzed by bioinformatics methods. The expression of KIF15 was examined in GC and paracarcinoma tissues from 41 patients to verify the analysis results. The relationship between KIF15 expression and clinical characteristics were also observed by bioinformatics methods. Kaplan-Meier survival analysis of 122 GC patients in our hospital was performed to explore the relationship between KIF15 expression levels and GC patients' prognosis. KIF15 was downregulated in GC cell lines AGS and SGC-7901 by transfecting a lentivirus-mediated shRNA plasmid targeting KIF15. In vitro, GC cell proliferation and apoptosis were detected by MTT assay, colony formation assay, and Annexin V-APC staining. In vivo, xenograft experiments were used to verify the in vitro results. Furthermore, Human Apoptosis Antibody Array kit was used to screen possible targets of KIF15 in GC cell lines.

Results

The bioinformatics results showed that KIF15 expression levels were higher in GC tissues than in normal tissues. IHC showed same results. High expression of KIF15 was statistical correlated with high age and early histologic stage. Kaplan-Meier curves indicated that high KIF15 expression predict poor prognosis in patients with GC. MTT assay and colony formation assay showed that KIF15 promote GC cell proliferation. Annexin V-APC staining found that KIF15 can inhibit GC cell apoptosis. Xenograft experiments reveal that downregulating KIF15 can inhibit GC tumor growth and promote GC apoptosis. Through detection of 43 anti-apoptotic proteins by the Human Apoptosis Antibody Array kit, it was confirmed that knocking down KIF15 can reduce seven anti-apoptotic proteins expression.

Conclusions

Taken together, our study revealed a critical role for KIF15 to inhibit GC cell apoptosis and promote GC cell proliferation. KIF15 may decrease anti-apoptotic proteins expression by regulating apoptosis pathways. High expression of KIF15 predicts a poor prognosis in patients with GC. KIF15 might be a novel prognostic biomarker and a therapeutic target for GC.

SUBMITTER: Ding L

PROVIDER: S-EPMC7160940 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Background: Dysregulation of prostate stem cell antigen (PSCA) has been implicated in human cancers. Studies have reported that PSCA expression is generally high in prostate cancer, which correlates with a worse survival. PSCA is also highly expressed in bladder, ovarian, and pancreatic cancers. However, PSCA is expressed at low levels in gastric, gallbladder and oesophageal cancers. At present, the clinical significance, expression pattern and biological function of PSCA in gastric cancer (GC) are still unclear. Methods: Previously, we used cDNA microarray as a screening tool to compare GC tissues with its matched normal gastric mucosa tissues (MNGT), and obtained the differentially expressed genes of the two tissue types. PSCA is one of the genes significantly down-regulated in GC tissues. In this study, we detected the expression of PSCA in GC tissues and MNGT by western-blot experiment and immunohistochemical staining (IHC). Then the relationship between the expression pattern of PSCA and the clinicopathological characteristics and survival in GC was analyzed. In order to further study the function of PSCA in GC, lentivirus was used to construct stable cell lines with knockdown and overexpression of PSCA gene. We used AGS and MKN45 cell lines for plasmid transfection. Colony formation assay, MTS and nude mice xenograft model were performed to investigate the effect of PSCA in GC. Results: Western-blot and IHC assays demonstrated that the expression of PSCA in GC tissues was significantly lower than that in the MNGT. PSCA expression in GC tissues was high in 252 (57.5%) and low in 186 (42.5%) of 438 patients. PSCA expression for MNGT was high in 273 (62.3%) and low in 165 (37.7%) of 438 patients. PSCA expression was significantly associated with T classification (P=0.024), N classification (P=0.018) and TNM stage (P=0.019) using χ2 test. The relationship between PSCA expression level and patient survival was analysed using Kaplan-Meier analysis and the log-rank test. Low levels of PSCA expression were significantly associated with a poorer OS than high expression levels of PSCA (P=0.011). In the COX regression analysis of OS, all 9 variables in the univariate analysis were significantly correlated with OS (P<0.05), while the variables found to be independently correlated with OS in the multivariate analysis were PSCA expression (P=0.036), age (P<0.001), gender (P=0.007), and TNM stage (P<0.001), respectively. Univariate and multivariate analyses showed that PSCA was an independent prognostic factor for OS in GC. In vitro MTS cell proliferation experiment and clonal formation experiment and in vivo nude mouse subcutaneous tumorigenesis experiment all proved that knockdown of PSCA gene can improve the proliferation ability of GC cells, while in vitro experiment proved that overexpression of PSCA can reduce the proliferation ability of GC cells.It was found that knockdown of PSCA gene can improve the proliferation ability of GC cells both in vitro and in vivo, while overexpression of PSCA can reduce the proliferation ability of GC cells in vitro. Conclusion: Our study showed that the expression of PSCA gene was decreased in GC, which was related to more advanced pathological stages. And the expression level of PSCA in GC was an independent good prognostic factor. PSCA gene had the function of inhibiting GC proliferation.

Project description:BackgroundGastric cancer is the second leading cause of cancer death and remains a major clinical challenge due to poor prognosis and limited treatment options. Long noncoding RNAs (lncRNAs) have emerged recently as major players in tumor biology and may be used for cancer diagnosis, prognosis, and potential therapeutic targets. Although downregulation of lncRNA GAS5 (Growth Arrest-Specific Transcript) in several cancers has been studied, its role in gastric cancer remains unknown. Our studies were designed to investigate the expression, biological role and clinical significance of GAS5 in gastric cancer.MethodsExpression of GAS5 was analyzed in 89 gastric cancer tissues and five gastric cancer cell lines by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Over-expression and RNA interference (RNAi) approaches were used to investigate the biological functions of GAS5. The effect of GAS5 on proliferation was evaluated by MTT and colony formation assays, and cell apoptosis was evaluated by hochest stainning. Gastric cancer cells transfected with pCDNA3.1 -GAS5 were injected into nude mice to study the effect of GAS5 on tumorigenesis in vivo. Protein levels of GAS5 targets were determined by western blot analysis. Differences between groups were tested for significance using Student's t-test (two-tailed).ResultsWe found that GAS5 expression was markedly downregulated in gastric cancer tissues, and associated with larger tumor size and advanced pathologic stage. Patients with low GAS5 expression level had poorer disease-free survival (DFS; P?=?0.001) and overall survival (OS; P?<?0.001) than those with high GAS5 expression. Further multivariable Cox regression analysis suggested that decreased GAS5 was an independent prognostic indicator for this disease (P?=?0.006, HR?=?0.412; 95%CI?=?2.218-0.766). Moreover, ectopic expression of GAS5 was demonstrated to decrease gastric cancer cell proliferation and induce apoptosis in vitro and in vivo, while downregulation of endogenous GAS5 could promote cell proliferation. Finally, we found that GAS5 could influence gastric cancer cells proliferation, partly via regulating E2F1 and P21 expression.ConclusionOur study presents that GAS5 is significantly downregulated in gastric cancer tissues and may represent a new marker of poor prognosis and a potential therapeutic target for gastric cancer intervention.