Background
Renal cell carcinoma (RCC) is the most common type of malignant renal cancer in adults, responsible for approximately 90–95% of the diagnosed cases [
1]. Approximatelys, 25–30% patients present metastasis at the time of diagnosis and 30% of the patients relapse after renal surgery [
2]. To date, surgery is the primary treatment for RCC, and the five-year survival rate is 65–90%; however, the outcome is considerably reduced in metastatic cases [
3]. RCC is relatively resistant to radiotherapy and chemotherapy with only a 4–5% response rate [
4,
5]. Some cases respond to immunotherapy with a 30% response rate [
6]. By 2013, with the advancement in targeted therapy, such as Sunitinib and Sorafenib, the average survival time was improved from 12 months to 22 months in patients with metastatic RCC [
7,
8]. However, the five-year overall survival for metastatic RCC remains <10% [
3].
ATP-binding cassette sub-family G member 2 (ABCG2) was first named as Breast Cancer Resistance Protein in the 1990s when it was discovered in MCF-7 breast cancer cell line co-selected for doxorubicin in the presence of verapamil [
9]. Following its discovery, ABCG2 was cloned, characterized, and added as the second member of the G subfamily of ABC transporters, as a semi-transport protein [
10]. ABCG2 has main effect on effluxing drugs at major physiological barriers, such as blood-brain, blood-testis and maternal-fetal barriers. Similar function of ABCG2 is validated in effluxing of xenobiotics at small intestine and kidney proximal tubule brush borders. ABCG2 transports a wide variety of substrates including several anticancer agents and is one of the most significant contributors to multidrug resistance in cancer cells [
10,
11].
Although ABCG2 has been studied in diverse fields, the precise function and effect in RCC are yet poorly understood [
12]. Due to the heterogeneous high expression pattern of ABCG2 in the kidney, researchers have speculated that this protein may be actively involved in drug resistance, leading to failure of chemotherapeutic treatments [
13,
14]. Clinically, whether ABCG2 expression could predict overall survival for RCC has not been well studied. Thus, we aimed to evaluate the correlation between ABCG2 expression and overall survival of patients with clear cell RCC managed by renal surgery.
Discussion
The prognosis of RCC is difficult to predict, and thus, a reliable biomarker is essential to guide the clinical management. RCC is a chemotherapy-resistant and radiotherapy-resistant malignant tumor, and the efficiency of immunotherapy for this type of cancer is also limited [
4‐
6]. The introduction of tyrosine-kinase inhibitors (TKIs) was considered as a breakthrough in the treatment of metastatic RCC [
16‐
18]. However, most metastatic RCC patients administrated with TKIs eventually yield to disease progression due to drug resistance [
19]. Several studies have been initiated in an attempt to discover reliable prognostic biomarkers to predict the overall survival of RCC patients. Bui et al. found that carbonic anhydrase IX and Ki67 are useful prognostic biomarkers for RCC that can improve the survival prediction and classification of renal cancer [
20]. Several other biomarkers, such as carbonic anhydrase 9, phosphatase, and tensin homologue deleted on chromosome 10, vimentin and p53, are also deemed to correlate with overall survival in RCC patients when in combined clinical parameters [
21]. Vermaat et al. also reported that serum amyloid α was a robust and independent prognosticator for overall survival in RCC patients [
22].
ABCG2 is a member of the ATP-binding cassette transporters and an ATP-dependent membrane protein predominantly expressed in the kidney [
23]. ABCG2 is highly expressed in cancer stem cells or side-population cells and may protect the cells by pumping out xenobiotics, detrimental metabolites of oxidative stress, and chemotherapeutic drugs [
23‐
25]. Recent studies have shown that ABCG2 has a vital role in the multidrug resistance of cancer cells and may influence the overall survival of tumor patients [
26,
27]. Yoh et al. reported that in advanced non-small cell lung cancer the response rate to chemotherapy in patients with ABCG2(−) tumors was 44%; however, in ABCG2(+) tumors, this rate decreased to 24%, accompanied by shorter overall survival than ABCG2(−) patients (
p = 0.004) [
28]. Interestingly, by utilizing the resources of TCGA database, we found that the mean expression level of ABCG2 in clear cell RCC is highest compared to all the other genitourinary and gynecologic tumors (Additional file
1: Fig. S1,
p < 0.001). So we speculate that ABCG2 minght be a crucial marker and regulator of clear cell RCC progression. Therefore, we evaluated the prognostic impact of ABCG2 expression on overall survival in clear cell RCC patients managed with renal surgery.
In the present study, we found that ABCG2 expression highly correlated with overall survival in patients with clear cell RCC. Overall survival is considered as the most valuable endpoint in terms of assessing the prognostic outcome in RCC patients, and the analysis of this parameter is a strength of the study. Survival curves (Fig.
2) showed a significant difference of overall survival among various degrees of expression of ABCG2,
p < 0.001. The stronger the ABCG2 expression, poorer the overall survival. The median overall survival was 93.2, 85.8, and 17.0 months in ABCG2(−), (+), and (++) subgroups, respectively. The five-year survival rate was 95%, 77%, and 27% in ABCG2(−), (+), and (++) subgroups, respectively. Based on the routine practice in our institution, interferon alpha or TKIs (Sorafenib or Sunitinib) were used as adjuvant therapeutics for patients with RCC after surgery; the efficacy of TKIs was reported as promising [
16‐
18]. However, in our study, 19 patients with metastatic RCC treated with TKIs showed a poor median overall survival.
Moreover, we analyzed the correlation of ABCG2 expression of these 19 patients with metastatic RCC. The median overall survival was 59.4 (IQR = 43.8–71.3) months in ABCG2(+) cohort and 16.8 (IQR = 14.5–19.8) months in ABCG2(++) cohort,
p < 0.001. The predisposition indicated that stronger the ABCG2 expression, poorer the prognosis. The correlation necessitates further studies. Interestingly, we found that ABCG2 expression significantly correlated with Fuhrman grade in renal cell carcinoma. This may be associated with the protein expression of the nucleus in malignant renal cell. Higher mRNA levels of ABCG2 gene was found in more malignant pancreatic cell [
29] and we need to elucidate this in further research in RCC. In the multivariate analysis, ABCG2 and Fuhrman grade were significant predictors for overall survival (
p < 0.001) when combined with the TNM status parameters.
Due to the limited knowledge and conditions, the mechanism of ABCG2 in RCC is not fully elucidated. Szakacs et al. reported that ABCG2 played a vital role in the multidrug resistance of cancer cells, thereby influencing the overall survival [
27]. Hypoxia is the key step in the development and progression of RCC and is mainly regulated by Von Hippel-Lindau, an important tumor suppressor gene. Besides, hypoxia is verified to affect the ATP-binding cassette transporter family, including ABCG2 [
30]. In the pancreatic cancer, there are significant correlations between mRNA levels of ABCG2 and clinical outcomes [
29]. Many researches are arising to detect the mechanism of ABCG2 in drug resistance in other malignancies, while, to our knowledge, studies regarding ABCG2 expression in RCC leading to therapeutic resistance are lacking. Till date, there is one report by Korenaga and colleagues indicating that the ABCG2 polymorphism (C421A) is a risk factor for developing non-papillary RCC [
31]. So it’s necessary to make further study in the field of ABCG2 effect on drug resistance in RCC. With the advent of efficient inhibitors of ABCG2, the combination strategies of targeted drugs and ABCG2 inhibitors might provide the promising therapeutic effect.
The detection ABCG2 expression by IHC staining is clinically valuable. Improved diagnostic techniques aimed at the selection of RCC patients with less expression of ABCG2 might result in more successful outcomes. For example, the RCC patients with highly expressed ABCG2 require more care after surgery and intensive follow-up. Moreover, IHC staining is a common and economical method to detect ABCG2 and could be widely used in clinical management. Therefore, ABCG2 could be utilized in most medical institutions.
While this study comprised a moderate size of patients with extended follow-up, it is also retrospective and derived from a single tertiary-care center, which could impact the generalizability. Another limitation is that the paraffin sections were preserved for a prolonged duration that may affect the IHC staining to a certain degree.
Acknowledgments
We sincerely appreciate the patients’ participation.