Shtabinskaya T.T., Kendysh E.N.

Grodno State Medical University, Belarus

Expression of angiogenic markers depending on the degree of colon adenocarcinoma differentiation

The main criterion of the degree of malignancy in pathologic diagnosis of colon adenocarcinomas according to the "International histological classification of tumors of the intestine" is the degree of differentiation of tumor cells. The relatively more favorable clinical form with a relatively satisfactory long-term results is typical for highly and moderately differentiated adenocarcinoma, and for low-grade, expressed different structural and cellular anaplasia, characterized by a more aggressive course and worse prognosis.

However, none of these factors can’t explain why, even with similar clinical and morphological traits, as well as the extent and nature of the surgery different survival. This fact was the promise to search for predictors that are based not only on clinical data and standard morphological studies, but also in the study of molecular and biological properties of the tumour [1, 2].

Purpose: to assess expression of angiogenic markers expression in colon adenocarcinoma depends on tumour differentiation.

Materials and methods: 72 cases of CRC were investigated. The cases were divided into 2 groups. The I group included 55 patients with low-grade adenocarcinoma (LGA) , in the II - 17 patients high-grade adenocarcinoma (HGA). The study was performed on paraffin sections using antibodies to VEGF, CD 105, MMP-2, Erk2, iNOS, TGF-β, NRP1, MMP-9 according to standard procedure. Quantitation of the level of expression was described in the previous article [3]. Statistical analysis was performed using STATISTICA 10.0 (SNAXAR207F394425FA-Q).

Results:

Levels positivity angiogenesis markers in LGA and HGA presented in Table 1.

Table 1 – Expression of CD105, VEGFA, NRP1, MMP-2, MMP-9, TGF-β, Erk2, iNOS, depending on the degree of tumor differentiation

Angiogenic markers		Grade		z	Р
		G1-2 (n=55)	G3 (n=17)
CD105	vascular endothelium	0,698 (0,477-0,774)	0,826 (0,654-0,893)	-1,830	0,067
VEGFА	Overall	0,982 (0,962-0,989)	0,781 (0,769-0,905)	3,592	<0,001
	parenchymal	0,992 (0,976-0,995)	0,719 (0,679-0,990)	3,216	0,001
	stromal	0,964 (0,946-0,988)	0,924 (0,809-0,986)	2,242	0,025
NRP1	Overall	0,735 (0,622-0,811)	0,574 (0,548-0,682)	1,742	0,008
	parenchymal	0,622 (0,547-0,700)	0,532 (0,510-0,544)	3,454	<0,001
	stromal	0,854 (0,758-0,920)	0,755 (0,699-0,791)	3,357	<0,001
MMP-2	Overall	0,052 (0,026-0,081)	0,023 (0,014-0,042)	2,489	0,013
	parenchymal	0,029 (0,013-0,068)	0,007 (0,004-0,011)	3,294	<0,001
	stromal	0,048 (0,029-0,093)	0,019 (0,012-0,051)	0,518	0,012
MMP-9	Overall	0,510 (0,382-0,638)	0,639 (0,336-0,777)	-1,365	0,172
	parenchymal	0,651 (0,462-0,762)	0,789 (0,520-0,899)	-1,542	0,122
	stromal	0,360 (0,245-0,466)	0,532 (0,371-0,651)	-2,257	0,023
TGF-β	Overall	0,493 (0,250-0,650)	0,910 (0,630-0,940)	-3,570	<0,001
	parenchymal	0,590 (0,412-0,847)	0,930 (0,748-0,960)	-2,915	0,004
	stromal	0,257 (0,106-0,406)	0,815 (0,590-0,850)	-4,048	<0,001
Erk2	Overall	0,019 (0,005-0,037)	0,009 (0,003-0,030)	0,916	0,359
	parenchymal	0,021 (0,004-0,040)	0,007 (0,003-0,029)	1,031	0,303
	stromal	0,003 (0,002-0,011)	0,003 (0,001-0,023)	0,463	0,644
iNOS	Overall	0,725 (0,660-0,820)	0,840 (0,650-0,900)	-1,338	0,181
	parenchymal	0,770 (0,620-0,860)	0,820 (0,680-0,970)	-1,373	0,169
	stromal	0,750 (0,670-0,850)	0,790 (0,660-0,840)	-0,464	0,643

According to the literature, TGF-β has an immunosuppressive effect, creates immune tolerance and prevent rejection of tumors, thereby contributing to its anaplasia; increases the expression of genes encoding MMPs are involved in the degradation of extracellular matrix and basement membrane. Our results showed that expression of TGF-β and MMP-9 in HGA was significantly higher than in KGA and median positivity MMP-2, whose expression, according to some literature data [4] is not related to the degree of tumor differentiation, below.

With the progression of the disease accumulates in the tumor cell clones negative for VEGF expression [5]. The VEGFA and NRP1 in LGA was significantly higher (p <0,05), than G3 tumors.

Also showed a trend toward reliability when compared CD105 positivity: in tumors with high grades is slightly larger (p = 0.067).

No statistically significant differences in the expression of iNOS and Erk2 depending on Grade tumors were found.

Conclusion: When cancer progresses with increasing Grade tumor increases the expression level of TGF-β, MMP-9 (p <0.01 and p = 0.023, respectively). And positivity VEGFA, NRP1 and MMP-2 is significantly higher in LGA compared to HGA (p <0,03; p <0,001; p <0,01; respectively).

Literature:

1. Nuclear thymidylate synthase expression in sporadic colorectal cancer depends on the site of the tumor/ V. Sulzyc-Bielicka [et al.] // Virchows Arch. – 2009. – Vol. 454. – №6. – P. 695-702.

2. Rectal and pouch recurrences after surgical treatment for familial adenomatous polyposis / F.G. Campos [et al.] // J Gastrointest Surg. – 2009. – Vol. 13. – №1. – P. 129-136.

3. Штабинская, Т.Т. Прогностическое значение уровня экспрессии фактора роста эндотелия сосудов в колоректальном раке / Т.Т. Штабинская [и др.] // Научно-практический журнал УО «Гродненский государственный медицинский университет». – 2015. - № 3(51). - С. 64-69.

4. Герштейн, Е.С., Матриксные металлопротеиназы 2, 3, 13 и их тканевой ингибитор 2-го типа в опухолях и плазме крови больных раком толстой кишки/ Е.С. Герштейн, Е.А. Короткова, В.В. Пророков, Н.Е. Кушлинский// Бюллетень экспериментальной биологии и медицины. – 2008. – Т. 145. – №3. - С. 337-341.

5. Карамышева, А.Ф. Анти-ангиогенная терапия: надежды и разочарования: Сигнальные системы, регулирующие ангиогенез опухоли, и их изменение при опухолевой прогрессии. – Lap Lambert Academic Publishing GmbH KG, 2013. – 72 с.