长链非编码RNA BANCR与非小细胞肺癌EGFR-TKI耐药的相关研究

目的：探讨长链非编码RNA BANCR与非小细胞肺癌表皮生长因子受体（EGFR）-酪氨酸激酶抑制剂（TKI）耐药之间的相关性。方法：选择EGFR19外显子区缺失突变的细胞株以及野生型细胞株进行研究，应用实时荧光定量PCR法（qRT-PCR）检测耐吉非替尼细胞株PC9/GR及亲本细胞株PC9中BANCR的表达差异，发现BANCR在PC9/GR细胞中显著低表达。在PC9/GR细胞中过表达BANCR，分别应用CCK8法、克隆形成实验、流式细胞术检测过表达后PC9/GR细胞对吉非替尼药物敏感性，细胞增殖能力，联合吉非替尼处理后细胞凋亡变化；Western blot检测过表达BANCR后对PC9/GR细胞上皮间质转化（EMT）的影响。结果：①吉非替尼的敏感细胞株的RNA BANCR表达水平较高，耐药细胞株的RNA BANCR表达水平较低（P<0.05）。②在PC9/GR细胞中过表达BANCR，相对于对照组，吉非替尼对PC9/GR细胞的半数抑制浓度（IC50）减低，前后差异具有统计学意义（P<0.05）。过表达BANCR后PC9/GR细胞增殖能力减弱，经吉非替尼处理后细胞凋亡增多（P<0.05）。③与空白对照组比较，Western blot结果显示过表达BANCR的PC9/GR细胞E-钙粘蛋白表达水平明显升高，N-钙粘蛋白水平明显降低。结论：长链非编码RNA BANCR可诱导细胞凋亡，抑制细胞增殖；过表达BANCR可以增加PC9/GR细胞对吉非替尼的敏感性，逆转其耐药性，这一作用机制可能是通过调控EMT过程来发挥作用的。

Research on the relationship between non-coding RNA BANCR in human non-small cell lung cancer with EGFR-TKI

Objective: To study the long chain noncoding RNA BANCR with non-small cell lung cancer with Epidermal growth factor receptor (Epidermal growth factor receptor, EGFR) - Tyrosine kinase inhibitors (Tyrosine kinase inhibitors, TKI) correlation between the resistance. Methods: choose EGFR19 exons area lack of mutant cell lines and wild type cell lines to study, application of real-time fluorescent quantitative PCR technique (qRT - PCR) to detect the treatment resistant cell line PC9/GR and parental cell lines in the PC9 BANCR expression differences, find BANCR in PC9/GR cells significantly lower expression. In PC9/GR cells expressing BANCR, were determined by CCK8 method, the clone formation experiment, flow cytometry to detect PC9/GR cells after expressing the treatment for drug sensitivity, cell proliferation, apoptosis changes after the treatment the joint treatment. Western blot detection after expressing BANCR PC9/GR cells the expression of EMT changes. The treatment results: (1) The sensitive cell lines of RNA BANCR express high levels and drug resistance cell line of the low level of RNA BANCR expression (P < 0.05).(2) In the PC9/GR cells expressing BANCR, compared with the control group, the treatment for, for the PC9/GR half inhibitory concentration (IC50) of cells to reduce, before and after the differences statistically significant (P< 0.05). The proliferation of PC9/GR cells decreased after overexpression of BANCR, and apoptosis increased after treatment with gefitinib (P < 0.05). (3)Compared with the blank control group, Western blot showed that the E-cadherin expression level of PC9/GR cells overexpressing BANCR was significantly increased, while the N-cadherin level was significantly decreased. Conclusion: Long-chain non coding RNA BANCR can induce apoptosis and inhibit cell proliferation; overexpression of BANCR can increase the sensitivity of pC9/GR cells to gefitinib and reverse their drug resistance, which may play a role by regulating the EMT process.