Yonsei Med J.  2018 Nov;59(9):1041-1048. 10.3349/ymj.2018.59.9.1041.

Heat Shock Factor 1 Predicts Poor Prognosis of Gastric Cancer

• Kim SJ
• Lee SC
• Kang HG
• Gim J
• Lee KH
• Lee SH
• Chun KH

• KMID: 2422488
• DOI: http://doi.org/10.3349/ymj.2018.59.9.1041

Abstract

PURPOSE
Heat shock factor 1 (HSF1) is a key regulator of the heat shock response and plays an important role in various cancers. However, the role of HSF1 in gastric cancer is still unknown. The present study evaluated the function of HSF1 and related mechanisms in gastric cancer.
MATERIALS AND METHODS
The expression levels of HSF1 in normal and gastric cancer tissues were compared using cDNA microarray data from the NCBI Gene Expression Omnibus (GEO) dataset. The proliferation of gastric cancer cells was analyzed using the WST assay. Transwell migration and invasion assays were used to evaluate the migration and invasion abilities of gastric cancer cells. Protein levels of HSF1 were analyzed using immunohistochemical staining of tissue microarrays from patients with gastric cancer.
RESULTS
HSF1 expression was significantly higher in gastric cancer tissue than in normal tissue. Knockdown of HSF1 reduced the proliferation, migration, and invasion of gastric cancer cells, while HSF1 overexpression promoted proliferation, migration, and invasion of gastric cancer cells. Furthermore, HSF1 promoted the proliferation of gastric cancer cells in vivo. In Kaplan-Meier analysis, high levels of HSF1 were associated with poor prognosis for patients with gastric cancer (p=0.028).
CONCLUSION
HSF1 may be closely associated with the proliferation and motility of gastric cancer cells and poor prognosis of patients with gastric cancer. Accordingly, HSF1 could serve as a prognostic marker for gastric cancer.

Keyword

Heat shock factor 1 (HSF1); gastric cancer; prognostic marker

MeSH Terms

Dataset
Gene Expression
Heat-Shock Response
Hot Temperature*
Humans
Kaplan-Meier Estimate
Oligonucleotide Array Sequence Analysis
Prognosis*
Shock*
Stomach Neoplasms*

Figure

• 

  Fig. 1 Evaluation of the correlation between HSF1 expression and survival rate of patients with gastric cancer using public databases. (A–C) Expression levels of HSF1 mRNA in tumor and normal tissues from the GEO database. Public datasets are presented as a scatter diagram: (A) GSE13861, (B) GSE13195, and (C) GSE30727. p values were calculated using Student's t-test (A and B: *p<0.001; C, p=0.061). (D) Kaplan-Meier survival plots demonstrate the poor prognostic effect of HSF1 upregulation, which was correlated with worse overall survival, in patients with gastric cancer (probe 1: 202344_at, n=876; probe 2: 213756_s_at, n=876). HSF1, heat shock factor 1; HR, hazard ratio.

• 

  Fig. 2 High expression of HSF1 associated with poor survival in patients with gastric cancer. (A) In situ expression of HSF1 in normal and tumor tissues from patients with intestinal and diffuse types of gastric cancer was detected by immunohistochemistry and hematoxylin and eosin staining (magnification: ×200). (B) Comparative analysis of HSF1 expression in normal and tumor tissues from patients with gastric cancer based on staining intensity. p values were calculated using Student's t-test and significant differences are indicated by * (*p<0.001). (C) Kaplan-Meier analysis of survival curves for patients with gastric cancer according to the expression of HSF1 in tumor tissues (n=54). HSF1 high-expression group (score 3) had a much lower survival rate at 125 months than the HSF1 low-expression group (score 1 or 2). HSF1, heat shock factor 1.

• 

  Fig. 3 Downregulation of HSF1 expression reduces proliferation, migration, and invasion of gastric cancer cells. AGS and MKN28 cells were transfected with a scrambled siRNA (scRNA) or two small-interfering RNAs (siRNAs) specific for HSF1 (siRNA #1 and #2). (A) HSF1 protein expression was detected by Western blot analysis. β-actin was used as a loading control. (B) WST assay was performed to detect cell viability. (C and D) Transwell assays to evaluate (C) migration and (D) invasion of cells (×200). The histogram is represented as mean±SEM (n=3). p values were calculated using ANOVA and statistically significant differences are indicated as * (*p<0.001). HSF1, heat shock factor 1.

• 

  Fig. 4 Overexpression of HSF1 promotes proliferation, migration, and invasion of gastric cancer cells. Overexpression of HSF1 in AGS and MKN28 cells was achieved by transfection with an empty vector (pcDNA_EV) or HSF1 overexpression vector (pcDNA_HSF1). (A) Expressions of HSF1 mRNA and protein were detected by Western blot analysis. β-actin was used as a loading control. (B) WST assay was performed to detect cell viability. (C and D) Transwell migration assays to evaluate (C) migration and (D) invasion activity of cells (×200). Data are represented as mean±SEM (n=3). p values were calculated using Student's t-test (*p<0.001, †p=0.001). HSF1, heat shock factor 1.

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