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. 2014 Jul;35(7):6311-7.
doi: 10.1007/s13277-014-1819-y. Epub 2014 Mar 19.

Overexpression of HCC1/CAPERα may play a role in lung cancer carcinogenesis

Yurong Chai  1 , Xinxin Liu, Liping Dai, Yang Li, Mei Liu, Jian-Ying Zhang
Affiliations

Overexpression of HCC1/CAPERα may play a role in lung cancer carcinogenesis

Yurong Chai et al. Tumour Biol. 2014 Jul.

Abstract

HCC1/CAPERα is considered to be a novel human tumor-associated antigen, and the tumor-specific immunity of HCC1/CAPERα has been reported in several types of cancer. However, there was very limited evidence indicating its function in tumorigenesis. In the present study, to elucidate the roles and underlying molecular mechanism of HCC1/CAPERα in lung cancer, we examined the expression of HCC1/CAPERα in human non-small cell lung cancer (NSCLC) cell line and NSCLC tissue microarray (TMA). Immunohistochemistry with TMA was performed to detect HCC1/CAPERα expression in NSCLC and adjacent lung tissues. NSCLC cell line constitutively transfected by pcDNA3.1-HCC1/CAPERα, and empty pcDNA3.1 vector were used. These cells were analyzed by Western blot, MTT, immunofluorescence, wound healing assay, and transwell assays. It was found that HCC1/CAPERα was mainly localized in the nucleus of the lung cancer cells and overexpression of HCC1/CAPERα may promote lung cancer cells proliferation and increase cells migration. The frequency of HCC1/CAPERα expression in NSCLC tissues was significantly higher than that in adjacent and normal tissues (P < 0.01). Our data suggest that overexpression of HCC1/CAPERα may increase the proliferation and migration of NSCLC cells, and HCC1/CAPERα could be a promising biomarker for lung cancer.

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Conflict of interest statement

Conflicts of interest None

Figures

Fig. 1
Fig. 1
Expression of HCC1/CAPERα in tumor cells. a Expression of HCC1/CAPERα in six tumor cell lines were analyzed by Western blotting. The polyclonal anti-HCC1/CAPERα antibody was used as a probe. H460 and H1299: lung cancer, AGS: gastric cancer, HepG2 and SUN449: liver cancer, SKBR3: breast cancer. β-Actin was used as loading control. b, c Expression of HCC1/CAPERα in H1299 cells were analyzed by Western blotting (b) and immunofluorescence staining (c). H1299 cells were transfected with empty vector pcDNA3.1 (vector), pcDNA3.1-HCC1.3 (HCC1.3), or pcDNA3.1-HCC1.4 (HCC1.4). It showed that HCC1/CAPERα was overexpressed in H1299-HCC1.4 cells. HCC1/CAPERα demonstrated stronger nuclear immunofluorescence staining pattern in H1299 cells transfected with pcDNA3.1-HCC1.4 and also found expressed in the perinuclear cytoplasm of dividing H1299 cells (c). The arrows indicate the diving cells
Fig. 2
Fig. 2
Proliferative activity of H1299 cells. MTT assay was used to estimate the proliferation of H1299 cells after transfected at different time points. OD: Optical density. Data shown are means±SD of at least three independent experiments. It showed that there was higher proliferation activity in H1299-HCC1.4 cells. Asterisk: designated statistical significant, P<0.05
Fig. 3
Fig. 3
Migration capability of H1299 cells. After being transfected, H1299 cells were subjected to transwell migration assay (a, c) and wound healing assay (b). a The representative fields on membrane. Migrated cells on the bottom membrane of transwell inserts were stained with crystal violet (magnification, ×10). b The transfected H1299 cells at once after applying stripes as control (0 h) and observed again after 24 h. The H1299-HCC1.4 cells nearly formed 100 % confluent monolayer after 24 h incubation. c Migrated cells were stained with fluorescence dye: calcein AM. The fluorescence data of migrated cells were collected by fluorescence plate reader. Data shown were means±SD of at least three independent experiments. It showed that the H1299-HCC1.4 cells had the significantly higher level of migration activity compared with H1299-vector cells. Asterisk statistically different to vector, P<0.05
Fig. 4
Fig. 4
Expression of HCC1/CAPERα in normal, adjacent lung, and NSCLC tissues by IHC. Tissue microarray slide was stained with polyclonal anti-HCC1/CAPERα antibody to detect the expression of HCC1/CAPERα. One representative normal lung tissue (a) and a representative adjacent lung tissue (b) were negatively stained; c one positive control prostate cancer tissue; d, e, and f representative NSCLC grades I, II, and III tissues were positively stained (magnification, ×400). The NSCLC tissue with higher grade was intensively staining. The arrows indicate positive nuclear staining
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