Department of Biotechnology, School of Bioengineering, College of Engineering and Technology, Faculty of Engineering and Technology, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, 603203, India.
Clin Transl Oncol. 2024 Jul;26(7):1630-1638. doi: 10.1007/s12094-023-03380-6. Epub 2024 Feb 3.
The current challenge in clinical cancer treatment is chemoresistance. Colon cells have inherently higher xenobiotic transporters expression and hence can attain resistance rapidly. Increased levels of TGF-β2 expression in patients have been attributed to cancer progression, aggressiveness, and resistance. To investigate resistance progression, we treated doxorubicin (dox) to HT-29 colon adenocarcinoma cells in the presence or absence of TGF-β2 ligand.
After 1, 3-, and 7-day treatment, we investigated cell proliferation, viability, and cytotoxicity by MTT, trypan blue staining, and lactate dehydrogenase enzyme release. The mechanism of cell death was elucidated by hoechst33342 and propidium iodide dual staining and apoptosis assay. The development of resistance was detected by rhodamine123 efflux and P-glycoprotein (P-gp)/MDR1 antibody staining through fluorimetry and flow cytometry. The colony formation ability of the cells was also elucidated.
Inhibition of cell proliferation was noted after day 1, while a significant reduction in viability and a significant increase in lactate dehydrogenase release was detected after day 3. Reduction of intracellular rhodamine123 levels was detected after day 3 and was significantly lower in dox with TGF-β2 treatment compared to dox alone. Increased surface P-gp levels after days 3 and 7 were observed in the treated groups. Hoechst33342/propidium iodide staining and apoptosis assay indicated non-apoptotic cell death. The cells treated with TGF-β2 had higher colony formation ability.
TGF-β2 expression might play a significant role in the development of chemoresistance to doxorubicin in Duke's type B colon adenocarcinoma cell line, HT-29.
目前临床癌症治疗的挑战是化疗耐药性。结肠细胞固有地具有更高的外源性化合物转运体表达，因此可以迅速获得耐药性。患者中 TGF-β2 表达水平的增加归因于癌症进展、侵袭性和耐药性。为了研究耐药性进展，我们在用或不用 TGF-β2 配体的情况下，用阿霉素（dox）处理 HT-29 结肠腺癌细胞。
在 1、3 和 7 天治疗后，我们通过 MTT、台盼蓝染色和乳酸脱氢酶酶释放来研究细胞增殖、活力和细胞毒性。通过 hoechst33342 和碘化丙啶双重染色和凋亡测定阐明细胞死亡的机制。通过 rhodamine123 外排和 P-糖蛋白（P-gp）/MDR1 抗体染色通过荧光法和流式细胞术检测耐药性的发展。还阐明了细胞的集落形成能力。
第 1 天观察到细胞增殖抑制，第 3 天观察到活力显著降低和乳酸脱氢酶释放显著增加。第 3 天检测到细胞内 rhodamine123 水平降低，与单独 dox 相比，dox 与 TGF-β2 处理后显着降低。在处理组中，第 3 天和第 7 天观察到表面 P-gp 水平增加。Hoechst33342/碘化丙啶染色和凋亡测定表明非凋亡性细胞死亡。用 TGF-β2 处理的细胞具有更高的集落形成能力。
TGF-β2 表达可能在 Duke 型 B 结肠腺癌细胞系 HT-29 中对阿霉素的化疗耐药性的发展中起重要作用。
