TRPM7 is involved in acid-induced necrotic cell death in a manner sensitive to progesterone in human cervical cancer cells
TRPM7 is involved in acid-induced necrotic cell death in a manner sensitive to progesterone in human cervical cancer cells
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DOI：
10.14814/phy2.14157
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发表时间：
2019-07-01
影响因子：
2.5
通讯作者：
Okada, Yasunobu
中科院分区：
文献类型：
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作者：
Numata, Tomohiro;Sato-Numata, Kaori;Okada, Yasunobu
由于阴道内pH呈强酸性，因此研究酸中毒对宫颈癌细胞的影响非常重要。最近，在强酸中毒的反应中，人宫颈癌HeLa细胞在显示出由Cl-内流诱导的持续细胞肿胀后显示出坏死。由于阳离子内流应伴随着Cl-内流，以驱动水内流引起细胞肿胀，我们在这里研究了酸毒阳离子电导的本质。RT-PCR和Western blotting检测mRNA/蛋白表达。膜片钳技术测量离子电流。细胞计数/活力和比色法分别用于评估增殖率和caspase 3/7活性。细胞体积和大小分别通过电子尺寸和视频显微镜测量来测量。酸暴露增强了TRPM 7活性，TRPM 7在HeLa细胞中内源表达，在HEK 293 T细胞中外源过表达。TRPM 7的基因沉默消除了酸诱导的细胞肿胀和坏死，但却诱导了HeLa细胞中凋亡半胱天冬酶3/7的激活。孔电荷中和D1054 A突变体的过表达抑制了HEK 293 T细胞中酸增强的阳离子电流、酸诱导的细胞肿胀和酸毒性坏死。令人惊讶地发现，Progressive治疗抑制HeLa细胞中TRPM 7的分子和功能表达以及细胞增殖。此外，在胆甾酮处理的细胞中，酸暴露不会诱导持续的细胞肿胀，然后坏死，但诱导持续的细胞皱缩和凋亡性细胞死亡。这些结果表明，在人宫颈癌细胞中，TRPM 7基本上参与酸毒性坏死细胞死亡，而黄体酮抑制TRPM 7表达，从而通过切换到细胞凋亡来抑制酸毒性坏死。
Because intravaginal pH is strongly acidic, it is important to investigate the effects of acidosis on cervical cancer cells. Recently, in response to strong acidosis, human cervical cancer HeLa cells were shown to exhibit necrosis after showing persistent cell swelling induced by Cl- influx. Since cation influx should be accompanied with Cl- influx to drive water inflow causing cell swelling, we here studied on the nature of acidotoxic cation conductance. The mRNA/protein expression was assessed by RT-PCR and Western blotting. Ionic currents were measured by patch-clamping techniques. Cell counting/viability and colorimetric assays were applied to assess proliferation rate and caspase 3/7 activity, respectively. Cell volume and size were measured by electronic sizing and video-microscopic measurements, respectively. Acid exposure enhanced TRPM7 activity endogenously expressed in HeLa cells and exogenously overexpressed in HEK293T cells. Gene silencing of TRPM7 abolished acid-induced cell swelling and necrosis but rather induced activation of apoptotic caspase 3/7 in HeLa cells. Overexpression with the pore charge-neutralizing D1054A mutant suppressed acid-enhanced cation currents, acid-induced cell swelling, and acidotoxic necrosis in HEK293T cells. Progesterone treatment was surprisingly found to suppress molecular and functional expression of TRPM7 and cell proliferation in HeLa cells. Furthermore, in the progesterone-treated cells, acid exposure did not induce persistent cell swelling followed by necrosis but induced persistent cell shrinkage and apoptotic cell death. These results indicate that in the human cervical cancer cells, TRPM7 is essentially involved in acidotoxic necrotic cell death, and progesterone inhibits TRPM7 expression thereby inhibiting acidotoxic necrosis by switching to apoptosis.