Abstract

Present study analyze the possible mutations in mitochondrial encoded genes TRNL1 and ND1, the association of mtDNA copy number in PCOS women with and without diabetic mothers was evaluated. Blood samples were collected from all the individual participants of study and detailed anthropometric and demographic variables were collected. Sequence of each amplified product of RNA gene TRNL1 was analyzed in comparison with the updated Cambridge Sequence. The PCR amplified products of ND1 were digested using Hae III and TaqI and were run in electrophoresis in 4% agarose gel electrophoresis. Relative quantification of mtDNA was compared with unique nuclear encoded gene based on the ratio of mtDNA to unique nuclear encoded gene using Real Time PCR. The group was effectively and closely similar for age and BMI. Aged varied between 24–25 years and 50–51 years for daughters and mother respectively. Results identified a five different type of mutation in study sample, among which only one was seen in non PCOS group, rest other four were found in both daughters and mother of PCOS group. All these nucleotide point mutation were verified by sequence analysis of both strands and appeared to be heteroplasmy Of the 36 PCOS samples analyzed one sample showed the G3316A mutation in the homoplasmy state. Nonsynonymous mutation leads to amino acid change from Alanine to Threonine. Analysis of the copy number in mothers revealed that mothers of PCOS women too had less copy number when compared to the mothers of Non-PCOS. Mtcopy number was significantly reduced in PCOS women with diabetic mother when compared to the PCOS women with non-diabetic mother and also the Mtcopy number was less in PCOS women of non-diabetic mother with diabetic family history. Though evidence is not confirmatory to say that PCOS is a phenotypic feature of MIDs. This study on mtDNA variants reported with small number of sample in association with PCOS by a single centre can be justified with more number of samples in multicentre studies. To explore more of hereditary PCOS, whole exome (genome) sequencing is suggested.

中文翻译：

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多囊卵巢综合征（PCOS）患者的线粒体DNA突变和ND1基因拷贝数

摘要

本研究分析了线粒体编码基因TRNL1和ND1的可能突变，评估了有或没有糖尿病母亲的PCOS妇女中mtDNA拷贝数的关联。从研究的所有个体参与者中采集血液样本，并收集详细的人体测量学和人口统计学变量。将RNA基因TRNL1的每个扩增产物的序列与更新的剑桥序列进行比较。使用Hae III和TaqI消化ND1的PCR扩增产物，并在4％琼脂糖凝胶电泳中进行电泳。使用实时PCR，根据mtDNA与唯一核编码基因的比率，将mtDNA的相对定量与唯一核编码基因进行比较。该组在年龄和BMI方面有效且紧密相似。女儿和母亲的年龄分别在24-25岁和50-51岁之间。结果确定了研究样本中的五种不同类型的突变，其中在非PCOS组中仅见一种突变，其余四种在PCOS组的女儿和母亲中均发现。所有这些核苷酸点突变均通过两条链的序列分析得到证实，并且似乎是异质的。在所分析的36个PCOS样品中，一个样品显示G3316A突变处于同质状态。非同义突变导致氨基酸从丙氨酸变为苏氨酸。对母亲的拷贝数进行分析后发现，与非PCOS的母亲相比，PCOS妇女的母亲的拷贝数也较少。与没有糖尿病母亲的PCOS妇女相比，患有糖尿病母亲的PCOS妇女的Mtcopy数目显着减少，并且有糖尿病家族史的非糖尿病母亲的PCOS妇女的Mtcopy数目也较少。尽管没有证据表明PCOS是MID的表型特征。单个中心报告的少量样本与PCOS相关的mtDNA变体的这项研究可以在多中心研究中用更多的样本来证明。为了探索更多的遗传性PCOS，建议进行全外显子组（基因组）测序。单个中心报告的少量样本与PCOS相关的mtDNA变体的这项研究可以在多中心研究中用更多的样本来证明。为了探索更多的遗传性PCOS，建议进行全外显子组（基因组）测序。单个中心报告的少量样本与PCOS相关的mtDNA变体的这项研究可以在多中心研究中用更多的样本来证明。为了探索更多的遗传性PCOS，建议进行全外显子组（基因组）测序。