Co-evolution of tumor-associated macrophages and tumor neo-vessels during cervical cancer invasion

Shuting Jiang, Yuehong Yang, Min Fang, Xianglang Li, Xiuxue Yuan, Jingping Yuan, Shuting Jiang, Yuehong Yang, Min Fang, Xianglang Li, Xiuxue Yuan, Jingping Yuan

Abstract

Considering the crucial significance of the tumor microenvironment in cancer development and progression, the present study aimed to investigate the changes in macrophages and angiogenesis during the cervical cancer (CC) progression process from chronic cervicitis to cervical intraepithelial neoplasia grades I-III (CIN I-III) to CC. This investigation included quantitative analysis and assessment of the spatial associations between tumor-associated macrophages (TAMs) and tumor neo-vessels. The conventional immunohistochemistry staining technique was used to detect cluster of differentiation (CD)68 and CD105 biomarker expression for TAMs and tumor neo-vessels, respectively. In addition, with the assistance of quantum dot (QD)-based two-component in situ imaging technology, the expression of the TAMs and tumor neo-vessels could be observed simultaneously. The quantitative analysis and co-evolution of the TAMs and tumor neo-vessels could then be processed. During the progression process from chronic cervicitis to cervical CIN I-III, and ultimately to invasive CC, the expression of the macrophages and neo-vessels in the tumor microenvironment increased synchronously. According to the quantitative analysis results, the median value of the TAM density was higher in the CC group (5,540.14) than in the CIN I-III group (2,502.17) and the chronic cervicitis group (1,403.31), with statistical significance in all three groups (P<0.001, for between-group comparisons). The number of neo-vessels was also much higher in the CC group (n=27) than in the CIN I-III group (n=17) or the chronic cervicitis group (n=6.5), with statistical significance in all three groups (P<0.001, for between-group comparisons). These findings demonstrated the great significance and close association of TAMs and tumor angiogenesis during CC development and progression. Thus, QDs-based in situ and simultaneous imaging of key cancer molecules may provide insights with regard to the biology of cancer invasion.

Keywords: cervival cancer; quantum dots; tumor neo-vessels; tumor-associated macrophages.

Figures

Figure 1.
Figure 1.
Establishment of QD-based molecular imaging of cervical cancer tissue. (Aa) Staining of TAMs by IHC at ×200 magnification, and the expression of TAMs marked by CD68 with QD-525 (green color) at (Ab) ×200 and (Ac) ×400 magnification. (Ba) Staining of neo-vessels by IHC at ×200 magnification, and the expression of tumor neo-vessels marked by CD105 with QD-585 (yellow color) at (Bb) ×200 and (Bc) ×400 magnification. TAMs, tumor-associated macrophages; IHC, immunohistochemistry; QD, quantum dot; CD, cluster of differentiation.
Figure 2.
Figure 2.
Dynamic changes in the tumor microenvironment during CC progression. (Aa) H&E staining, (Ab) TAM staining with QDs-525 and (Ac) tumor neo-vessel staining with QD-585, in normal tissue. (Ba) H&E staining, (Bb) TAM staining with QDs-525 and (Bc) tumor neo-vessel staining with QD-585, in CC in situ tissue. (Ca) H&E staining, (Cb) TAM staining with QD-525 and (Cc) tumor neo-vessel staining with QD-585, in well-differentiated CC. (Da) H&E staining, (Db) TAM staining with QD-525 and (Dc) tumor neo-vessel staining with QD-585, in poorly-differentiated CC. Aa-Ac, Ba and Bb, Ca-Cc and Da-Dc: Magnification, ×200; and Bc: Magnfication, ×400. TAMs, tumor-associated macrophages; QD, quantum dot; CC, cervical cancer; H&E, hematoxylin and eosin.
Figure 3.
Figure 3.
QD-based TAM and tumor neo-vessel staining in situ simultaneously. H&E staining of (Aa) chronic cervicitis tissue, (Ba) CC in situ and (Ca) invasive CC tissue at ×200 magnification, and TAMs and tumor neo-vessels simultaneously stained with QD-525 and QD-585 in chronic cervicitis tissue at (Ab) ×200 and (Ac) ×400 magnification, in CC in situ at (Bb) ×200 and (Bc) ×400 magnification, and in invasive CC tissue at (Cb) ×200 and (Cc) ×400 magnification. TAMs, tumor-associated macrophages; QD, quantum dot; CC, cervical cancer; H&E, hematoxylin and eosin.

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