Decreased sensitivity of several anticancer drugs in TMEPAI knockout triple-negative breast cancer cells

Wardhani, Bantari Wisynu Kusuma; Puteri, Meidi Utami; Watanabe, Yuji; Louisa, Melva; Setiabudy, Rianto; Kato, Mitsuyasu

doi:10.13181/mji.v28i2.2687

Cited by 3 publications

(4 citation statements)

References 23 publications

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“… 33 , 34 A previous study by Wardhani et al reported that without TGF-β stimulation, the response of TNBC Hs578T to cytotoxic drugs would be reversed. 35 In our study, doxorubicin retained its cytotoxic effect result in both TMEPAI positive and negative cells. Nonetheless, TMEPAI knock-out cells showed a more potent result.…”

Section: Discussionsupporting

confidence: 57%

TGF-β-Induced TMEPAI Promotes Epithelial–Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration

Wardhani

Louisa

Watanabe

et al. 2021

BCTT

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Introduction Epithelial–mesenchymal transition (EMT) and overexpression of drug efflux transporters have been reported to cause doxorubicin resistance. Our previous study indicated that TMEPAI (transmembrane prostate androgen-induced protein) attenuated doxorubicin sensitivity in triple-negative breast cancer cells. However, how TMEPAI contributes to doxorubicin resistance in TNBC remains unclear. Thus, the present study aimed to elucidate the mechanism of TMEPAI in doxorubicin resistance in triple-negative breast cancer cells. Methods We used BT549, triple-negative cells wild type (WT), and BT549 TMEPAI knock-out. Both cells were treated with TGF-β 2 ng/mL for 24 hours, followed by TGF-β 2 ng/mL and doxorubicin 12.9 nM for another 24 hours. Afterward, the cells were harvested and counted. Cells were further lysed and used for RT-PCR and Western blot analysis. We determined the expression levels of proliferation, apoptosis, EMT markers, and drug efflux transporters. Additionally, we investigated the expressions of PI3K as well as SMAD3 and AKT phosphorylation. Results TNBC cells were shown to be less sensitive to doxorubicin in the presence of TMEPAI. TMEPAI was shown to alleviate the mRNA expressions of apoptosis markers: Bax, Bcl2, Caspase-3, and Caspase-9. Our results indicated that the presence of TMEPAI greatly amplifies EMT and increases drug efflux transporter expressions after doxorubicin treatment. Furthermore, our findings demonstrated that TMEPAI reduced the action of doxorubicin in inhibiting SMAD3 phosphorylation. TMEPAI was also shown to modify the effect of doxorubicin by reducing PI3K expressions and Akt phosphorylation in triple-negative breast cancer cells. Conclusion Our findings indicate that TMEPAI promotes EMT and drug efflux transporters at least in part by shifting doxorubicin action from SMAD3 phosphorylation reduction to PI3K/AKT inhibition in triple-negative breast cancer cells.

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Section: Discussionsupporting

confidence: 57%

TGF-β-Induced TMEPAI Promotes Epithelial–Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration

Wardhani

Louisa

Watanabe

et al. 2021

BCTT

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“…In the absence of TGF-β, wild-type TNBC cells were more sensitive to cytotoxic than the KO-cells, followed by a decrease in drug efflux transporters, P-gp, and BCRP expressions. 38 Therefore, we suggest that the effect of TMEPAI on the change in cell response to anticancer is highly dependent on TGF-β.…”

Section: Discussionmentioning

confidence: 89%

“…18,36,37 In our previous study, we reported that in TNBC-KO, a higher dose of doxorubicin, paclitaxel, and cisplatin as compared to wild type, which is in contrast with our present result. 38 TMEPAI-KO cells showed decreased sensitivity to several chemotherapy drugs. In this system, TGF-β was not used to induce TMEPAI.…”

Section: Discussionmentioning

confidence: 97%

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<p>TGF-β-Induced TMEPAI Attenuates the Response of Triple-Negative Breast Cancer Cells to Doxorubicin and Paclitaxel</p>

Wardhani¹,

Puteri²,

Watanabe³

et al. 2020

JEP

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Purpose: Triple-negative breast cancer (TNBC) is a refractory type of breast cancer with poor prognosis and limited choice for treatment. Previous studies had shown that TNBC has high expressions of transmembrane prostate androgen-induced protein (TMEPAI). TMEPAI was known to be induced by TGF-β/Smad signaling and have tumorigenic functions that converting TGF-β from tumor suppressor to tumor promoter and inducing epithelialmesenchymal transition (EMT). Therefore, we aimed to define the role of TMEPAI in triple-negative breast cancer cells treatment using several anti-cancers in the presence of TGF-β. Methods: TMEPAI-knock out (KO) was carried out in a triple-negative breast cancer cell, BT549. TMEPAI editing was developed using the CRISPR-Cas9 system using two combinations of sgRNA to remove exon 4 of the TMEPAI gene entirely. Genotyping and proteomic analysis were performed to check the establishment of the TMEPAI-KO cells. Wild type (WT) and KO cells were used to determine inhibitory concentration 50% (IC 50) of several anti-cancers: doxorubicin, cisplatin, paclitaxel, and bicalutamide in the presence of TGF-β treatment. Results: KO cells were successfully established by completely removing the TMEPAI gene, which was proven in genomic and proteomic analysis. Further, in TMEPAI-KO cells, we found a significant reduction of IC 50 for doxorubicin and paclitaxel, and minimal effects were seen for cisplatin and bicalutamide. Our findings suggest that TGF-β-induced TMEPAI attenuates the response of TNBC to doxorubicin and paclitaxel, but not to cisplatin and bicalutamide. Conclusion: TGF-β induced TMEPAI contributes to the reduced response of TNBC treatment to doxorubicin and paclitaxel, but minimal on cisplatin and bicalutamide. Further study is needed to confirm our findings in other growth factor-induced cells, as well as in in vivo model.

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CRISPR technology: A versatile tool to model, screen, and reverse drug resistance in cancer

Shirani-Bidabadi

Tabatabaee

Tavazohi

et al. 2023

European Journal of Cell Biology

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Decreased sensitivity of several anticancer drugs in TMEPAI knockout triple-negative breast cancer cells

Cited by 3 publications

References 23 publications

TGF-β-Induced TMEPAI Promotes Epithelial–Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration

TGF-β-Induced TMEPAI Promotes Epithelial–Mesenchymal Transition in Doxorubicin-Treated Triple-Negative Breast Cancer Cells via SMAD3 and PI3K/AKT Pathway Alteration

<p>TGF-β-Induced TMEPAI Attenuates the Response of Triple-Negative Breast Cancer Cells to Doxorubicin and Paclitaxel</p>

CRISPR technology: A versatile tool to model, screen, and reverse drug resistance in cancer

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