Figure 2

Overexpression of HCCR-1 induces the proliferation of PANC-1 cells. A) PANC-1 cells were transfected with control empty vector and HCCR-1 gene. B) HCCR-1 accelerated the proliferation of PANC-1 cells by 1.5 fold compared to control. The measurement of proliferation rate was performed by MTT assay. Values represent the mean ± SEM, Groups with annotations of H1 and H2 are significantly different with Vector, P < 0.01, two-way ANOVA. C) PANC-1 cells were transfected with HCCR-1 siRNA and control empty vector. D) Knock-down of HCCR-1 decreased the proliferation of PANC-1 cells MTT assay. Values represent the mean ± SEM, Groups with annotations of siRNA were significantly different with Vector, P < 0.01, two-way ANOVA. E) The invasiveness of HCCR-1 siRNA and vector transfectants were assessed by incubating cells in transwell plates(Costar) with polycarbonate filters for 24 h using 10% FBS as a chemoattractant. Cells invaded through the matrigel and filtered to the lower surface were fixed with 4% neutral-buffered formalin and stained with Giemsa. The number of invasion cells was significantly lower in HCCR-1 siRNA transfectants (24.4 ± 9.9) than in vector control(49.1 ± 15.4), *P < 0.01. Each experiment was repeated for three times. F) Immunohistochemical staining of pancreatic cancer tissues on a tissue chip. Strong straining of HCCR-1 in a pancreatic carcinoma(the upper two pictures). Expression of HCCR-1 in a paraneoplastic tissue(the two pictures below). The staining was graded according to Allred score system. G) The expression of HCCR-1 was analyzed by performing western blotting in the same tissues used in immunohistochemical staining. Lanes 1 to 3, paraneoplastic tissues; lanes4 to 7, pancreatic tumor tissues. Tubulin was used as a loading control.