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SCIENCE CHINA Life Sciences, Volume 60 , Issue 5 : 528-535(2017) https://doi.org/10.1007/s11427-016-0226-7

PSG9 promotes angiogenesis by stimulating VEGFA production and is associated with poor prognosis in hepatocellular carcinoma

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  • ReceivedOct 6, 2016
  • AcceptedNov 4, 2016
  • PublishedJan 4, 2017

Abstract


Funding

National Natural Science Foundation of China(81172035)

National High Technology Research and Development Program of China(2012AA020206)

National Basic Research Program of China(2014CBA02004)


Acknowledgment

This work was supported by the National Natural Science Foundation of China (81172035), National High Technology Research and Development Program of China (2012AA020206), and National Basic Research Program of China (2014CBA02004).


Interest statement

The author(s) declare that they have no conflict of interest.


References

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  • Figure 1

    PSG9 is preferentially expressed in tumors and frequently up-regulated in HCC. A, mRNA levels of PSG9 in paired HCC tissue samples. Total RNA was extracted from paired liver tissue samples, and PSG9 mRNA levels were detected by qRT-PCR, with 18S RNA as an internal control. B, The mRNA expression of PSG9 was correlated with that of VEGFA. P=0.015; R=0.501.

  • Figure 2

    Ectopic expression of PSG9 in SMMC7721 cells enhanced VEGFA expression. A, The luciferase activity of the VEGFA-promoter plasmid was enhanced by transfection with the pcPSG9 plasmid. *, P<0.05. B and C, Up-regulated PSG9 increased VEGFA mRNA levels in SMMC7721 cells. SMMC7721 cells were transiently transfected with pcDNA3.1 and pcPSG9. Total RNA was analyzed by qRT-PCR, and protein lysates were analyzed by western blot with antibodies against PSG9, VEGFA, and β-actin at 24 and 48 h. *, P<0.05.

  • Figure 3

    Role of PSG9 in proliferation, vascular remodeling and neoangiogenesis via the modulation of VEGFA. A, Up-regulation of PSG9 significantly accelerated growth, while down-regulated PSG9 inhibited the growth of SMMC7721 cells. B, The Matrigel assay was used to determine the ability of HUVECs to form capillaries when cultured with conditioned media from different cells. A closely knit network of capillary-like structures was found in the HUVECs incubated with the medium from the SMMC7721-PSG9 cells. C, The density of the capillary-like network was measured, and the results showed that more neoangiogenesis occurred in HUVECs incubated with conditioned medium from SMMC7721-PSG9 cells than in those from SMMC7721, SMMC7721-vector, or SMMC7721-PSG9 shRNA cells. *, P<0.05; **, P>0.05. D, The Matrigel plug model was developed, followed by the establishment of Matrigel sponges of SMMC7721, SMMC7721-vector, SMMC7721-PSG9, and SMMC7721-PSG9 shRNA cells. E, The Matrigel was excised and photographed. The intensity of red hemoglobin was measured.

  • Figure 4

    The PSG9 protein levels and Kaplan-Meier estimates of overall survival in cancer patients. A, Scatter plots of the PSG9 levels in plasma examined by ELISA. Mean levels of the proteins are represented by horizontal lines. B, PSG9 status in HCC plasma is associated with the disease-free survival of patients with HCC. P=0.042. C, PSG9 status in HCC plasma is associated with the overall survival of patients with HCC. P=0.002.

  • Table 1   Independent influence on overall survival of HCC patients, assessed with Cox’s proportional hazards model analysis

    Variables

    Univariate analysis

    Multivariate analysis

    HR (95% CI)

    P

    HR (95% CI)

    P

    PSG9 high vs. low*

    3.089 (1.473−6.480)

    0.003

    3.114 (1.434−6.762)

    0.004

    Vascular invasion (yes vs. no)

    1.279 (0.626−2.611)

    0.499

    TNM stage (I−II vs. III−IV)

    1.576 (0.779−3.189)

    0.206

    Tumor size (≤3 cm vs. >3 cm)

    0.395 (0.163−0.958)

    0.040

    0.379 (0.145−0.988)

    0.047

    Differentiation (L vs. M/H)**

    1.872 (1.023−3.425)

    0.042

    1.808 (0.979−3.340)

    0.058

    Tumor number (single vs. multiple)

    0.567 (0.234−1.373)

    0.209

    Cirrhosis (yes vs. no)

    0.887 (0.422−1.863)

    0.751

    HBV infection (yes vs. no)

    0.640 (0.225−1.819)

    0.403

    *, PSG9 levels in peripheral plasma of HCC patients examined by enzyme-linked immunosorbance assay, high, ≥30.44 ng mL−1; low, <30.44 ng mL−1. **, H, high differentiation; M, moderate differentiation; L, low differentiation, HR, Hazard ratio; 95% CI, 95% confidence interval.

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