ercc1 rrm1 top1 top2a tyms tubb3 gstp1 and brca1 … · received: 17/04/2020; accepted: 09/05/2020...

JBUON 2020; 25(4): 1728-1736ISSN: 1107-0625, online ISSN: 2241-6293 • www.jbuon.comEmail: [email protected]

ORIGINAL ARTICLE

Corresponding author: Matvey M. Tsyganov, PhD. Department of Experimental Oncology, Cancer Research Center, Tonsk Na-tional Research Medical Center, Russian Academy of Sciences. Kooperativny street no.7, Tomsk 634050, Russian Federation. Email: [email protected] Received: 17/04/2020; Accepted: 09/05/2020

Prognostic significance of ERCC1, RRM1, TOP1, TOP2A, TYMS, TUBB3, GSTP1 and BRCA1 mRNA expressions in patients with non-small-cell lung cancer receiving a platinum-based chemotherapyMatvey M. Tsyganov1, Evgeny O. Rodionov1, Alina M. Pevzner1,2, Marina K. Ibragimova1, Sergey V. Miller1, Olga V. Cheremisina1, Irina G. Frolova1, Sergey A. Tuzikov1, Nikolai V. Litviakov1

1Department of Experimental Oncology, Cancer Research Institute, Tomsk National Research Medical Center, Russian Academy of Sciences; 5, Kooperativny Street, 634050, Tomsk, Russian Federation. 2Tomsk State University, Tomsk, Lenina Ave 36, 634050, Russian Federation.

Summary

Purpose: Currently, more researchers are attracted by the possibility of assessing the sensitivity of a lung tumor to certain chemotherapy drugs and their personalized choice based on an assessment of this sensitivity. The purpose of this study was to explore the prognostic significance of the level of 8 chemosensitivity genes’ expression in patients with non-small cell lung cancer (NSCLC).

Methods: The study included 59 patients with NSCLC IIB–IIIA stage. RNA was isolated from the surgical material of the tumor and normal lung tissue. The expression level of 8 chemosensitivity genes BRCA1, RRM1, ERCC1, TOP1, TOP2a, TUBB3, TYMS, GSTP1 was evaluated using RT-PCR.

Results: Cases with higher metastasis-free survival rates showed a significantly low level of expression of the ERCC1,

BRCA1, GSTP1 genes (p=0.0004, p=0.01, p=0.01). In addi-tion, analysis of the overall survival revealed that the high-est rates were achieved in patients with overexpression of the ERCC1 gene. The overall survival in these patients was 86%, versus 55% in the other group and the differences were statistically significant (p=0.002). No statistically significant differences were found for the remaining genes.

Conclusion: Thus, a comprehensive assessment of the che-mosensitivity genes expression is important not only from the point of view of understanding the heterogeneity and complexity in the field of molecular biology of NSCLC, but also for a more accurate prognosis and course of the disease.

Key words: adjuvant chemotherapy, chemosensitivity genes, gene expression, non-small cell lung cancer, non-metastatic survival, overall survival

Introduction

Platinum-based chemotherapy plus different cytotoxic agents such as vinorelbine, gemcitabine, taxanes, etc., is the standard for the treatment of patients with non-small cell lung cancer (NSCLC) [1]. Systemic chemotherapy can be used in neoad-juvant and/or adjuvant settings. Early researches have shown that the effectiveness of neoadjuvant

chemotherapy (NAC) is highly variable in patients with NSCLC [2-4]. In a report by Pisters et al [5] the authors reported about 12% (9 of 73) of patients who achieved a complete response. Roth et al [6] showed a lack of efficiency of NAC in 28 patients while Ro-sell et al [7] had only 3% (1 of 30) patients with an objective response to treatment. In general, the

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Significance of mRNA of some genes in lung cancer 1729

JBUON 2020; 25(4): 1729

average frequency of complete regression described in these studies of NAC is about 4% (range 0-16). In the vast majority of patients with stage III NSCLC (about 35-50% of all patients with NSCLC) the progression of the process is due to the devel-opment of distant metastases [8]. Additionally, us-ing the NAC and/or surgical treatment this doesn’t lead to improved survival of patients and the treat-ment of such patients remains a challenge [9]. A study by Adjuvant Navelbine International Trial-ist Association (ANITA) randomized patients with stage IB, II, or IIIA into two groups. In the first group, AC was performed with the vinorelbine/cisplatin scheme in 4 courses, and in the second group patients were put under observation alone. Chemotherapy was found to increase the median survival (65.7 versus 43.7 months; RR 0.80; 95% CI 0.66-0.96, p=0.02) [10]. Currently, the main pro-gress in the treatment of lung cancer is associated with the use of targeted drugs - tyrosine kinase inhibitors of 1-3 generations and drugs against chimeric genes [11]. However, there are signifi-cant limitations for these drugs: firstly, they can only be used in a limited sample of patients with driver mutations, secondly, resistance to targeted therapy always develops and in the future it is necessary to switch to chemotherapy [12]. In this regard, many researchers are attracted by the pos-sibility of assessing the sensitivity of the tumor to certain chemotherapy drugs and their personalized choice based on the assessment of this sensitivity. There is an opinion that this is a new, promising area of treatment for patients with lung cancer [13]. One approach to personalizing chemotherapy for NSCLC is to evaluate the expression of chemosen-sitivity genes, which determines the sensitivity of tumor cells to individual chemotherapy drugs and the purpose of a chemotherapy regimen depending on the level of their expression [14]. Gene excision repair product (ERCC1) is a structurally specific en-donuclease involved in DNA repair. Clinicopatho-logical studies have shown that high expression of ERCC1 is associated with resistance to platinum-based chemotherapy [15] and overexpression of the BRCA1 gene is also associated with low cisplatin ef-fectiveness and low rates of relapse-free and overall survival [16]. Low RRM1 expression is a predictor of high survival in the treatment of gemicitabine chemotherapy [17]. For the enzyme thymidylate synthase (TYMS gene), it was found that its high expression is statistically significantly correlated with sensitivity to gemcitabine [18]. Thymidylate synthase is involved in the formation of de novo thymidylate, the precursor of thymidine triphos-phate, the nucleotide required for DNA synthesis. Overexpression of β3-tubulin (TUBβ3) is associated

with resistance to docetaxel and paclitaxel [19]. For doxorubicin, the most important group of the gene expression of the topoisomerase is topoisomer-ase 1 (TOP1) and 2α (TOP2α) [20]. These enzymes change the DNA topology, catalyze the unwinding of DNA supercoils, and the breaking and crosslink-ing of nucleic acid molecules. For the personalize AC treatment it is necessary to know not only the relationship between the level of chemosensitivity gene expression with a direct response to treat-ment, but also the relationship with the outcome of the disease. Recent studies have shown that TOP2a can be a good prognostic factor in the treatment of patients with NSCLC [21]. Patients with higher levels of expression had lower relapse-free survival compared to TOP2α-negative patients and multi-variate analysis showed that the level of TOP2α is an independent prognostic factor [22]. In this study, we analyzed the association of the chemosensitivity genes expression in the tumor tissue of NSCLC with the main clinical and patho-logical characteristics of the disease, as well as the indicators of overall and non-metastatic survival.

Methods

The study group

This study included 59 patients with NSCLC stage IIB–IIIA, central or peripheral localization with a mor-phologically verified diagnosis, who were treated at the clinic of the Oncology Research Institute of Tomsk Sci-entific Research Center. The study was conducted ac-cording to the ethical principles suggested in the 2013 Declaration of Helsinki [23] and approved by the Ethical Committee of the Cancer Research Institute. Signed in-formed consent was obtained from all participants. All patients received 2 courses of NAC with oral vinorelbine (25 mg/m2, day 1 and 8)/carboplatin AUC 6 (day 2) with a 3-week interval. Assessment of tumor response to NAC was based on the standard definitions according to re-sponse evaluation criteria in solid tumors (RECIST). All patients then underwent pneumonectomy or lobectomy followed by 3 courses of AC with “platinum doublets” according to the following schemes: vinorelbine 25 mg/m2 (1st and 8th days)/carboplatin AUC 6 (on the 2nd day); doxorubicin 50 mg/m2/carboplatin AUC 6 (on the 2nd day); gemcitabine 1250 mg/m2 (1st and 8th day)/carbo-platin AUC 6 (on the 2nd day); carboplatin AUC6 (on day 2)/paclitaxel 175 mg/m2. The interval between chemo-therapy courses was 3 weeks. Chemotherapy was dis-continued if there was evidence of disease progression, intolerability, or major toxicities. If during the NAC a good response was achieved, then the patients continued the vinorelbine-carboplatin regimen as AC. If there was a poor response to NAC or there were pronounced side effects, the chemotherapy regimen changed. The main clinical and morphological parameters of the patients are presented in Table 1.

Significance of mRNA of some genes in lung cancer1730

JBUON 2020; 25(4): 1730

Surgical material after chemotherapy (tumor tissue, unchanged lung tissue, ~30-60 mm3) was used as the test material. The tissues were placed in RNAlater (Ambion, USA), incubated for 24 h at room temperature and stored in –80ºС until RNA extraction.

RNA extraction

RNA was extracted from 59 matched normal and tumor tissues using RNeasy Plus mini Kit (Qiagen, Ger-many) according to the manufacturer’s instructions.

The RNA concentration and purity were assessed using NanoDrop 2000 instrument (Thermo Scientific, Waltham, USA). The concentration varied between 100 to 500 ng/µl and А260/А280 and А260/А230 ratios were 1.85-2.05 and 1.80-2.08, respectively. The RNA integrity was assessed using TapeStation instrument and R6K Screen-Tape kit (Agilent Technologies, Santa Clara, USA). The RIN values were 6.6-9.2. The RNA was reversely tran-scribed into cDNA using RevertAid™ kit (Thermo Fisher Scientific, Waltham, Lithuania) according to the manu-facturer’s instructions.

Expression profiling of the chemosensitivity genes

Expression profiling of the BRCA1, RRM1, ERCC1, TOP1, TOP2a, TUBB3, TYMS, and GSTP1 genes was car-ried out using quantitative real-time PCR (qRT-PCR) using custom fluorescent labelled probes and Rotor-Gene-6000 instrument (Corbett Research, Sydney, Aus-tralia). qRT-PCR was performed in triplicate in a volume of 15 µl containing 250 lM dNTPs (Sibenzyme, Novosi-birsk, Russia), 300 nM forward and reverse primers, 200

Clinical morphological parameters n (%)

Age, years

≤50 6 (10.2)

>50 53 (89.8)

Gender

Male 49 (81.1)

Female 10 (16.9)

Size of tumor

Т1 2 (3.4)

Т2 6 (10.2)

Т3 45 (76.3)

Т4 6 (10.2)

Lymph node metastasis

N0 15 (25.4)

N1 26 (44.1)

N2 18 (30.5)

Stage

IIB 14 (23.7)

IIIA 45 (76.3)

Clinical and anatomical location

Central 30 (50.8)

Peripheral 29 (49.2)

Histological type

Squamous 38 (64.4)

Adenocarcinoma 21 (35.6)

Effect of NAC

Complete regression 2 (3.4)

Partial regression 21 (35.6)

Stabilization 36 (61.0)

Type of surgery

Pneumonectomy 27 (45.8)

Lobectomy 32 (54.2)

AC regimens

Vinorelbine / carboplatin 36 (61.0)

Doxorubicin / carboplatin 4 (6.8)

Gemcitabine / carboplatin 16 (27,1)

Carboplatin / paclitaxel 3 (5.1)

Hematogeneous metastasis

Yes 17 (28.8)

No 42 (71.2)

Table 1. Μain clinical morphological parameters of pa-tients

Gene Amplicon (bp)

Sequence

GAPDH 124 bp F 5'-gccagccgagccacatc-3'

R 5'-ggcaacaatatccactttaccaga-3'

Probe 5'-cgcccaatacgaccaaatccg-3'

RRM1 94 bp F 5'-actaagcaccctgactatgctatcc-3'

R 5'-cttccatcacatcactgaacacttt-3'

Probe 5'-cagccaggatcgctgtctctaacttgca-3'

ERCC1 121 bp F 5'-ggcgacgtaattcccgacta-3'

R 5'-agttcttccccaggctctgc-3'

Probe 5'-accacaacctgcacccagactacatcca-3'

BRCA1 107 bp F 5'-acagctgtgtggtgcttctgtg-3'

R 5'-cattgtcctctgtccaggcatc-3'

Probe 5'-catcattcacccttggcacaggtgt-3'

TOP1 97 bp F 5'-ggcgagtgaatctaaggataatgaa -3'

R 5'- tggatatcttaaagggtacagcgaa -3'

Probe 5'-accattttcccatcatcctttgttctgagc -3'

TOP2A 75 bp F 5'-agtcgctttcagggttcttgag-3'

R 5'-tttcatttacaggctgcaatgg-3'

Probe 5'-cccttcacgaccgtcaccatgga-3'

TUBB3 71 bp F 5'-gggccaagttctgggaagtc-3'

R 5'-cgagtcgcccacgtagttg-3'

Probe 5'-atgagcatggcatcgaccccagc-3'

TYMS 91 bp F 5'-tctggaagggtgttttgga-3'

R 5'-tcccagattttcactccctt-3'

Probe 5'-tctttagcatttgtggatcccttga-3'

GSTP1 84 bp F 5'-ctggtggacatggtgaatgac-3'

R 5'-cttgcccgcctcatagttg-3'

Probe 5'-aggacctccgctgcaaatacatctc-3'

All Probes: FAM→BHQ1; bp - base pair, F - forward primer, R - reverse primer

Table 2. The sequence of primers and probe of genes


JBUON 2020; 25(4): 1731

nM probe, 2.5 mM MgCl2, 19 SE buffer (67 mM Tris–HCl pH 8.8 at 25 C, 16.6 mM (NH4)2SO4, 0.01% Tween-20), 2.5U Hot Start Taq polymerase (Sibenzyme, Novosi-birsk, Russia), and 50 ng of cDNA template. Samples were heated for 10 min at 95°C, followed by 40 cycles of amplification for 10 s at 95°C and 20 s at 60°C. Primer and probe (FAM-BHQ1) sequences were designed using Vector NTI Advance 11.5, Oligo 7.5, and the NCBI Nu-cleotide Database (https://www.ncbi.nlm.nih.gov/gene/) (Table 2). The mean expression level of each target gene was calculated for tumor tissue normalized to GAPDH (glyceraldehydes-3-phosphatedehydrogenase) and ACTB (actin beta). The average cycle threshold (Ct) was esti-mated for both the genes of interest and GAPDH and ACTB. Relative expression was evaluated using the Pfaffl method [24] (Table 2).

Statistics

Statistical analysis was performed using Statistica 8.0 software (StatSoft, Round Rock, USA). The arithme-tic mean value and standard error were calculated for each sample group. The expression level of the studied genes was divided by quartiles using basic statistics. Wilcoxon-Mann-Whitney test was used to assess the

differences between the studied groups. The compari-son of the frequencies in qualitative data was analyzed using the two-tailed Fisher’s exact test. The 95% confi-dence intervals (95% CIs) were calculated with the exact method. All p values were two-tailed and a p value of 0.05 was considered significant. To analyze the overall (OS) and non-metastatic survival, survival curves were constructed using the Kaplan-Meier method and the log-rank test was used to evaluate the differences in survival between groups.

Results and discussion

In a total group of 59 patients, distant me-tastases developed in 17 (29%) patients within 2-84 months from the date of diagnosis. One-year non-metastatic survival was 77.4%, 2-year 71.7%, 3-year 64.6%, 4-year 57.5; and 5-year 26.1%. To identify the relationship between the expression parameters of the studied genes with the clinical and pathological characteristics and patient sur-vival, we used the statistical characteristics of our

Figure 1. Non-metastatic survival (A, B, C) and overall survival (D) curves of 59 patients with non-small cell lung cancer depending on the level of mRNA expression of the ERCC1 gene, (log-rank test p=0.0004); BRCA1 (log-rank test p=0.01); GSTP1 (log-rank test p=0.03); ERCC1 (log-rank test p=0.002).


JBUON 2020; 25(4): 1732

sample in terms of gene expression. As a result, two analysis groups were obtained: patients with overexpression of genes (expression greater than 1, high expression) and the hypoexpression group (expression less than 1, low expression). As a result, at the first stage, the relationship of the expression of the studied chemosensitivity genes with the main clinical and pathological pa-rameters of the patients was evaluated (Table 3). T he expression levels of TYMS and TUBB3 were statistically significantly lower in the group of male patients (p=0.02 and p=0.03, respectively). It was also shown that high expression Top2A was more often observed in patients with stage IIB dis-ease (64.3%, 9/14 cases), compared with stage IIIA (31.1%, 14/45 cases), with p=0,03. The remaining clinical and morphological factors did not show a connection with the expression of the studied genes (Table 3). Using the Kaplan-Meier method, it was shown that with low expression of the ERCC1, BRCA1, and GSTP1 genes, there was higher metastatic-free survival rates compared to the group of patients with a high level of expression of these genes (Figure 1A, B, C). In particular, for the ERCC1 gene, it was found that the 4-year survival rate for the ERCC1 group <1 was 80% compared to 29% of the other group (log-rank test, p=0,0004), (Figure 1A). The same result was shown for the BRCA1 gene (Figure 1B). Non-metastatic survival rates were statistically significantly lower, i.e. 55% ver-sus 83% (with log-rank test p=0.01) in patients with a high level of expression of the studied genes. The GSTP1 gene is involved in the metabo-lism of xenobiotics and plays an important role in the detoxification of cells, including tumor cells, from the effects of chemotherapeutic drugs, in particular, platinum-based drugs [25,26]. Figure 1C shows the curves of non-metastatic survival rates depending on the level of expression of this gene. In the group of patients with a GSTP1 level of less than 1, the survival rate was 80% versus 54% in the group with GSTP1 expression of more than 1 (log-rank test, p=0.03). In addition, at the level of tendency, it was shown that the expression level of the RRM1 gene was not significantly associated with metastasis-free survival (p=0.1). No statistically significant differences were found for the remaining genes: TOP1 (p=0.83), Top2A (p=0.85), TUBB3 (p=0.41), TYMS (p=0.55). An analysis of OS found that the highest rates were achieved in patients with overexpression of the ERCC1 gene (Figure 1D). The OS in these pa-tients was 86% versus 55% in the opposite group, and the differences were statistically significant

(log-rank, p=0.002). Other genes showed no rela-tionship with OS: TOP1 (p=0.4), Top2A (p=0.84), TUBB3 (p=0.39), TYMS (p=0.52), GSTP1 (p=0.60), BRCA1 (p=0.73). As a result of this study, a relationship with the outcome of NSCLC was established for the three genes ERCC1, BRCA1 and GSTP1 which suggests the prospect of further studies of these genes as markers for personalized administration of AC to patients with NSCLC. In the resent study Sun et al it was shown that the high expression of ERCC1, RRM1, TUBB3 and TYMS connected with worse disease prognosis (p<0.001, p=0.001, p=0.001 and p=0.001). These genes are key factors affecting survival after sur-gical treatment in patients with NSCLC [27]. A lat-er study showed that only patients with low RRM1 expression had better progression-free survival rates (p<0.05) [28]. Interestingly, the results of meta-analyses indicate controversial effects of ERCC1 expres-sion on the prognosis of NSCLC. In particular, a 2010 meta-analysis found that the response to chemotherapy and metastatic survival rates were significantly higher in patients with low/nega-tive expression of ERCC1 (p<0.00001), compared with a group of patients with high expression of ERCC1 [29]. The level of ERCC1 mRNA was higher in tumors in patients with metastases (p=0.021) and higher in adenocarcinomas than squamous cell carcinomas (p=0.006), which had a more poor prognosis [30]. Another study also showed that relapse-free survival rates against the back-ground of ACT with platinum preparations were associated with a lack of expression of ERCC1 (HR, 2.166; 95% CI, 1.049-4.474; p=0.037) and BRCA1 (HR, 2.419; 95% CI , 1.127–5.193; p=0.023), but not with the expression level of RRM1 (HR, 0.568; 95% CI, 0.234–1.379; p=0.212) and TUBB3 (HR, 1.874; 95% CI, 0.448-7.842; p=0.39) [31]. However, OS rates were not associated with ERCC1 levels in the lung tumor of patients receiving surgical treatment with AC (OR=1.41; 95% CI, 0.93–2.12; p=0.106). Several authors as well [32] showed lack of association of β3-tubulin expression with sur-vival [33,34]. For the TYMS gene it was shown that this marker is weakly associated with OS and RFS [36], but there was a dependence on the chemotherapy scheme. Aberrant expression of this gene deter-mines low DFS with taxanes (p=0.012), with plati-num (p=0.003) and the combined taxane/platinum scheme (p=0.006). GSTP1 gene product is an en-zyme involved in the metabolism of chemothera-py drugs in tumor cells [35,36]. Basically, all stud-ies on the effect of this gene on the effectiveness


JBUON 2020; 25(4): 1733

Para

met

erER

CC

1>1

n (%

)ER

CC

1<1

n (%

)p-

leve

lB

RC

A1>

1n

(%)

BR

CA

1<1

n (%

)p-

leve

lG

STP

1>1

n (%

)G

STP

1<1

n (%

)p-

leve

lT

OP

1>1

n (%

)T

OP

1<1

n (%

)p-

leve

l

Age

≤50

1 (1

6.7)

5 (8

3.8)

12

(33.

3)4

(66.

7)0.

672

(33.

3)4

(66.

7)1

1 (1

6.7)

5 (8

3.3)

1

>50

11 (2

0.8)

42 (7

9.2)

26 (4

9.1)

27 (5

0.9)

18 (3

4.0)

35 (6

6.0)

8 (1

5.1)

45 (8

4.9)

Gen

der

Mal

e10

(20.

4)39

(79.

6)1

23 (4

6.9)

26 (5

3.1)

116

(32.

7)33

(67.

3)0.

726

(12.

2)43

(87.

8)0.

33

Fem

ale

2 (2

0.0)

8 (8

0.0)

5 (5

0.0)

5 (5

0.0)

4 (4

0.0)

6 (6

0.0)

3 (3

0.0)

7 (7

0.0)

Tum

or s

ize

Т1-

20

(0)

8 (1

00.0

)0.

182

(25.

0)6

(75.

0)0.

253

(37,

5)5

(62,

5)1

1 (1

2.5)

7 (8

7.5)

1

Т3-

412

(23.

5)39

(76.

5)26

(51.

0)25

(49.

0)17

(33.

3)34

(66.

7)8

(15.

7)43

(84.

3)

Lym

ph n

ode

met

asta

sis

N0

5 (3

3.3)

10 (6

6.7)

0.26

7 (4

6.7)

8 (5

3.3)

15

(33.

3)10

(66.

7)1

3 (2

0.0)

12 (8

0.0)

0.67

N1-

37

(15.

9)37

(84.

1)21

(47.

7)23

(52.

3)15

(34.

1)29

(65.

9)6

(13.

6)38

(86.

4)

Stag

ing

IIB

4 (2

8.6)

10 (7

1.4)

0.45

6 (4

2.9)

8 (5

7.1)

0.76

4 (2

8.6)

10 (7

1.4)

0.75

3 (2

1.4)

11 (7

8.6)

0.67

IIIA

8 (1

7.8)

37 (8

2.2)

22 (4

8.9)

23 (5

1.1)

16 (3

5.6)

29 (6

4.4)

6 (1

3.3)

39 (8

6.7)

Typ

e of

lung

can

cer

Cen

tral

4 (1

3.3)

26 (8

6.7)

0.20

14 (4

6.7)

16 (5

3.3)

18

(26.

7)22

(73.

3)0.

276

(20.

0)24

(80.

0)0.

47

Peri

pher

al8

(27.

6)21

(72.

4)14

(48.

3)15

(51.

7)12

(41.

4)17

(58.

6)3

(10.

3)26

(89.

7)

His

tolo

gy

Squa

mou

s8

(21.

1)30

(78.

9)1

15 (3

9.5)

23 (6

0.5)

0.11

12 (3

1.6)

26 (6

8.4)

0.77

5 (1

3.2)

33 (8

6.8)

0.70

Ade

noca

rcin

oma

4 (1

9.0)

17 (8

1.0)

13 (6

1.9)

8 (3

8.1)

8 (3

8.1)

13 (6

1.9)

4 (1

9.0)

17 (8

1.0)

Con

tinu

ed o

n th

e ne

xt p

age

Tabl

e 3.

Ass

ocia

tion

bet

wee

n m

RN

A e

xpre

ssio

n of

ER

CC

1, B

RC

A1,

GST

P1,

TO

P1,

Top

2A, R

RM

1, T

YMS,

TU

BB

3, a

nd c

lini

cal c

hara

cter

isti

cs


JBUON 2020; 25(4): 1734

Para

met

erTo

p2A

>1n

(%)

Top2

A<1

n (%

)p-

leve

lR

RM

1>1

n (%

)R

RM

1<1

n (%

)p-

leve

lT

YM

S>1

n (%

)T

YM

S<1

n (%

)p-

leve

lT

UB

B3>

1n

(%)

TU

BB

3<1

n (%

)p-

leve

l

Age

, yea

rs

≤50

2 (3

3.3)

4 (6

6.7)

12

(33.

3)4

(67.

7)1

0 (0

)6

(100

.0)

0.33

4 (6

6.7)

2 (3

3.3)

0.39

>50

21 (3

9.6)

32 (6

0.4)

17 (3

2.1)

36 (6

7.9)

12 (2

2.6)

41 (7

7.4)

23 (4

3.4)

30 (5

6.6)

Gen

der

Mal

e18

(36.

7)31

(63.

3)0.

4915

(30.

6)34

(69.

4)0.

717

(14.

3)42

(85.

7)0.

0219

(38.

8)30

(61.

2)0.

03

Fem

ale

5 (5

0.0)

5 (5

0.0)

4 (4

0.0)

6 (6

0.0)

5 (5

0.0)

5 (5

0.0)

8 (8

0.0)

2 (2

0.0)

Tum

or s

ize

Т1-

25

(62.

5)3

(37.

5)0.

242

(25.

0)6

(75.

0)0.

714

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418

(35.

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(64.

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(66.

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(15.

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(54.

9)

Lym

ph n

ode

met

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sis

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7 (4

6.7)

8 (5

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0.54

6 (4

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9 (6

0.0)

0.52

4 (2

6.7)

11 (7

3.3)

0.71

5 (3

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10 (6

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0.37

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316

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(63.

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5)8

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ing

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9 (6

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5 (3

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0.03

6 (4

2.9)

8 (5

7.1)

0.51

4 (2

8.6)

10 (7

1.4)

0.45

5 (3

5.7)

9 (6

4.3)

0.54

IIIA

14 (3

1.1)

31 (6

8.9)

13 (2

8.9)

32 (7

1.1)

8 (1

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37 (8

2.2)

22 (4

8.9)

23 (5

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lung

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tral

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18 (6

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7414

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Peri

pher

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1)8

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(55.

2)

His

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5813

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9)1

17 (4

4.7)

21 (5

5.3)

1

Ade

noca

rcin

oma

7 (3

3.3)

14 (6

7.7)

6 (2

8.6)

15 (7

1.4)

4 (1

9.0)

17 (8

1.0)

10 (4

7.6)

11 (5

2.4)

ER

CC

1: e

xcis

ion

repa

ir c

ross

-com

plem

enti

ng 1

; B

RC

A1:

bre

ast

canc

er 1

; G

STP

1, g

luta

thio

ne S

-tra

nsfe

rase

Pi

1; T

OP

1: D

NA

top

oiso

mer

ase

I; To

p2A

: D

NA

top

oiso

mer

ase

II a

lpha

; R

RM

1: r

ibon

ucle

otid

e re

duct

ase

subu

nit

M1;

TYM

S: t

hym

idyl

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synt

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; TU

BB

3: c

lass

III

β-t

ubul

in


JBUON 2020; 25(4): 1735

of treatment of NSCLC are focused on the study of its normal genetic variability [36,37]. Thus, the question of the prognostic signifi-cance of the level of chemosensitivity genes ex-pression and the possibility of their usage for personalized ACT is open. To be able to conduct meta-analyses, more research is needed in this direction on different samples. In addition, a pro-spective study with the appointment of ACT in terms of the expression of chemosensitivity genes has become an important link in assessing the prospects of using these markers for personalized treatment of NSCLC.

Conclusion

As a result of this study, it was found that the expression of the repair genes ERCC1, BRCA1, as well as the GSTP1 metabolism gene in patients with NSCLC can be predictive factors. Patients with low or no expression of ERCC1, BRCA1,

and GSTP1 had higher rates of OS and non-met-astatic survival with platinum-based adjuvant chemotherapy.

Funding

The reported study was funded by RFBR, pro-ject number 20-015-00023.

Ethical approval

All procedures performed in studies involv-ing human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Conflict of interests

The authors declare no conflict of interests.

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ercc1 rrm1 top1 top2a tyms tubb3 gstp1 and brca1 … · received: 17/04/2020; accepted: 09/05/2020...

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